常用引物信息列表
实验室常用缓冲液 常用引物序列汇总

实验常用试剂、缓冲液的配制方法Na2HPO4,2 mM KH2PO4 1 M Tris-HCl 、11M Tris-HCl □组份浓度□配制量□配制量1L 1L (pH7.4,7.6,8.0)□配置方法1. 称量下列试剂,置于1L烧杯中。
烧杯中。
□配置方法1. 称量121.1gTris置于1LNaCl 加入约800mL的去离子水,充分搅拌溶解。
8 g 2.KCl 0.2g3. 按下表量加入浓盐酸调节所需要的pH值。
Na2HPO4 1.42 g 浓值HCl pHKH2PO4 0.27g 7.4约70mL2. 向烧杯中加入约800 mL的去离子水,充分搅拌溶解。
7.6 约60mL3. 滴加HCl将pH42mL 8.0约值调节至7.4,然后加入去离子水将溶液定容至1L。
4. 将溶解定容至1L。
4. 高温高压灭菌后,室温保存。
5. 高温高压灭菌后,室温保存。
注意:上述PBS Buffer中无二价阳离子,如需要,可在配方中pH注意:应使溶液冷却至室温后再调定pH值,因为Tris溶液的补充1mM CaCl2和0.5 mM MgCl2。
pH值随温度的变化差很大,温度每升高1℃,溶液的值大约降低6、10 M醋酸铵0.03个单位。
□组份浓度10 M醋酸铵□配制量100mL 1.5 M Tris-HCl 2、1.5 M Tris-HCl □组份浓度□配置方法1. 称量77.1g醋酸铵置于100~配制量pH8.8 ()□1L 200 mL烧杯中,加入约30 mL的去离子水搅拌溶解。
1L1. □配置方法称取181.7gTris置于烧杯中。
2. 加入约800mL2.加去离子水将溶液定容至100mL。
的去离子水,充分搅拌溶解。
3.使用8.8pH3. 用浓盐酸调值至。
0.22μm滤膜过滤除菌。
4.密封瓶口于室温保存。
1L 4. 将溶液定容至5. 高温高压灭菌后,室温保存。
注意:醋酸铵受热易分解,所以不能高温高压灭菌。
7、Tris- HCl平衡苯酚□溶液的注意:应使溶液冷却至室温后再调定pH值,因为Tris配置方法1. 使用原料:大多数市售液化苯酚是清亮无色的,pH值大约无需重蒸馏℃,溶液的值随温度的变化差异很大,温度每升高pH1便可用于分子生物学实验。
常用载体测序引物列表

常⽤载体测序引物列表常⽤载体测序引物列表载体名称正向引物反向引物pACT T7 T3pACT2 GAL4 AD(5’AD) 3'AD/pACT2-R PACYCDUET-1(MCSII) DuetUP2 T7TERpAD/pl-DEST CMV-FpAD-Track pAD-Track-F ⽆pAS2-1 5’BD M13-26/48 PB42AD PB42ADF PB42ADR PBAD PBAD-F PBAD-R pBABE pBABE5’ pBABE3’ PBACPAK8 BAC1BAC2 pBK-CMV T7/M13-20 T3/M13-26 PBS(SK/KS)/M13- M13F/T7T3/M13R pBV220 PBV220F PBV220R pBI121需根据酶切位点确认需根据酶切位点确认 pCAMBIA 1301(1300)需根据酶切位点确认需根据酶切位点确认 pCAMBIA 2300 需根据酶切位点确认需根据酶切位点确认PCANTB5E S1/M13R S6pCAT3-enhancer pGL3+pcDNA3.0 pEGFP-N5/T7 SP6/BGHpcDNA3.1 pEGFP-N5/T7 BGHpcDNA4 T7/pEGFP-N5 BGHpcDNA6 T7/pEGFP-N5 BGHpcDNAII T7SP6 pCE2.1 M13FM13R pCEP4 pCMV5R/PCEP-F pEGFP-N5/EBV-R pCF-T M13FM13R pCIT7(17Base ) pCMV5R pCI-neo T7(17Base ) T3pCMS-EGFP T7T3 pCMV-3Tag-4A T3 /pFlag-CMV-F T7pCMV5 pCMV5F pCMV5R pCMV5-Flag pCMV5FpCMV5R pCMV-MYC/NUC pCMV5FpCMV5R PCMV-HA pCMV5FpCMV5R pCMV-SportSP6/M13RpCMV-Tag T3T7 pCMV-TNT T7/SP6pCMV5R pCR2.1-TOPO M13F/T7M13R pCR3.1 T7BGH pCS2 SP6T7PDNR-LIB M13F M13RpDonar M13F M13R pDONR201 pDONR-F pDONR-R pDONR221 M13F M13RpDrive T7 SP6 pDRIveR T7 SP6pDsRED1-C1 pDsRED-C1-F pECFP-C-3 pDsRed1-N1 pEGFP-N-5 RFP-NrevpDsRed-Monomer-N1 pEGFP-N-5 pDsRed1-NpDsRED2-C1 pDsRED-ex-C1-F pEGFP-N-3’pEGFP-N-5 pDsRed1-N pDSRED2-N1pDSRED-N1 pEGFP-N-5’ PDSRED-N-R pEASY-BLUNT M13F/T7 M13RTerpEASY-E1 T7 T7pEASY-T1 M13F/T7 M13RpEASY-T3 M13F/T7 M13R/SP6 pECFP-C pEGFP-C-5’ pECFP-C-3' pECFP-N1 pEGFP-N-5’ pEGFP-N-3’pEF/myc/ER pEF-F pCDNA3.1R pEGFP-C pEGFP-C-5’ pEGFP-C-3' pEGFP-N pEGFP-N-5’ pEGFP-N-3’ pENTR/D-TOPO M13F M13RTerpET-*(His) T7 T7pET22(a,b,c) T7 T7TerTerpET28,30,24,49 T7 T7TerpET32(a,b,c) T7/S.tag T7pET-42a (-b, -c)(+) Stag T7 TerpET50 T7/s.tag T7TERpET44 T7/s.tag T7Ter pEYFP-N1 pEGFP-N-5’ pEGFP-N-3’ PETBlue M13-20 PETBlueDOWN PETDUET-1(MCSI) PBRrevBam DuetDOMN1 PETDUET-1(MCSII) DuetUP2 T7TERpEYFP-C1 pEGFP-C-5’ pEGFP-C-3' pEYFP-N1 pEGFP-N-5’ pEGFP-N-3’ pFastBac pFastBac-F pFastBac-R pFastBac HT_A,B,C pFastBac-PH pECFP-C-3 pFastBac1 pFastBac-PH pECFP-C-3 pFastBac Dual (MCSI) pFastBac-PH pECFP-C-3 pFLAG-CMV CMV30/pFLAG-CMV-F CMV-24/pFLAG-CMV-Rp3xFlag-CMV-7.1 CMV-F/CMV24 CMV30pGAD424 5’AD 3'ADpGADGH 5’AD 3'ADpGADT7-Rec 5’AD/T7 3'ADpGAPZa-A/B/C a-FACTOR 3'AOXforward 3'AOXpGAPZ-A/B/C pGAPpGBKT7 T7 3'BDpGEM-T(-Easy) M13F/T7 SP6/M13RpGEX-(*)T pGEX-5’ pGEX-3’pGL2 pCMV5R PGL3-Pgl3-BASIC pGL3+ PGL3-pGL3-Enhancer pGL3+ PGL3-PinPoint TM PinPoint Primer SP6pIRES PIRES-F/T7 T3/PIRES-RpIRES2-EGFP pIRES2-EGFP.P5’ pIRES2-EGFP.P3’ pLenti6/v5-Dest pEGFP-N-5’PLEXA PLEXA-F PLEXA-R PLNCX PLNCX-F PLNCX-R pLXSN pLXSN-F pLXSN-RPjc1-tac T7pJet.1.21.Blunt T7primer M13F(-47) pMAL-c2E MalEP3/PMAL-C2X-R pMAL-C2x P5/PMAL-C2X-Fprimer M13F(-47) pMAL-p2X MalEPMD18-T M13R(-48) M13F(-47) PMD19-T M13F(-47) M13R(-48) pMIR-report M13FPPC86 PPC86-F PPC86-R3’AOXpPIC9K 5’AOX/a-factor3'AOX pPICZa 5'AOX/PPICZa-FpPROEXHTA M13R(-48)pQE30or40 pQE30+ pQE30- pRECEIVER CMV-FpRSET T7 T7TER PRSFDUET-1(MCSI) DuetUP1 DuetDOWN1 PRSFDUET-1(MCSII) DuetUP2 T7TER pShuttle-CMV pShuttle-CMV-F pShuttle-CMV-R PSILENCE-1.0-U6 T7 T3pSilencer3.1-H1 hygro M13F(-47) 3.0revpSILENCEV2.0-U6 T7 2.0revpSK01-T M13F M13RpSK-CMV T7 T3Psos H1PSP64/65 SP6pSP72 T7 SP6T7/M13RpSport1 SP6/M13F(-47)pSTBlue-1 T7 SP6pSUPER T7 T3pT7Blue(R) T7 M13F(-47)pTA2 M13F/T7 T3/M13R/T7 M13RpT-Adv M13FpTARGET TM pTarget-F/T7 arget-RPTHIOHISA,B,C TRX-FORWARD TRX-REVERSE pTO-T7 T7 T7TERPBV220-/PTRC99C-R pTRC99a-c PQE30+/PTRC99C-F pTriPLEx2 PT5/5'TriplEx2 T7pTz57r/t M13+ M13-pTWIN-1 T7 T7TERpUC18(19)/118(119) M13F(-47) M13R(-48)M13F(-47)/M13F pUC57 M13R(-48)/M13RpUCm-T M13F/T7 M13RpVAX1 T7 BGHT3/M13R/M13R(-48) pWSK29 T7/M13F/M13F(-47)pXT7 T7 SP6pYES2 T7 pYes2.R。
测序通用引物列表

测序通⽤引物列表测序通⽤引物列表载体名称上游引物(5'primer)下游引物(3'primer)P3*FLAG-CMV CMV-30(825-854) CMV-24(1129-1152) pAc5.1/V5-His(_A,_B,_C) pAc5-5 BGH rev pAcG2T pGEX-5 not availablepACT T7EEV T3/EBVrevpACT2 GAL4 ADfor pGAL4-ADrvpACT2 p17110 p12584pACYC184(BamHI-site) PBRforBam PBRrevBampACYCDuet-1(MCSI) ACYCDuetUP1 Primer DuetDOWN1pACYCDuet-1(MCSII) DuetUp2 T7-Terminator pAdEasy-1 not available not availablepAdTrack-CMV not available not available pAS2-1 GAL4 Bdfor/P24990 pGAL4-BDrv/P31430pB42AD pB42ADF PB42ADRpBABE pBMN_5'pBacPAK8 BAC1 BAC2pBAD pBAD-F pBAD-R pBAD/HisABC pBAD-5'pBBR1MCS T3/M13rev T7/M13forpBD-GAL4 pBD-GAL4-F PBD-GAL4-RpBI121 35S NOSpBIND GAL4-Bdfor T3,EBVrevpBK-CMV M13F/T7 M13R/T3 pBluescriptIIKS(+/-) T3/M13rev T7/M13for(优先⽤M13F)pBluescriptIISK(+/-) T3/M13rev T7/M13forpBluescriptKS(+/-) T3/M13rev T7/M13forpBluescriptSK(+/-) T3/M13rev T7/M13forpBR322(BamHI-site) PBRforBam PBRrevBampBR322(EcoRI/HindIII-sites) pBRforEco pBRrevHind pBR325(BamHI-site) PBRforBam PBRrevBampBR329(BamHI-site) PBRforBam PBRrevBam pBridge(MCSI) GAL4-Bdfor 3'BDpBridge(MCSII) not available not available pBS-T II T3/M13rev T7/M13for pBS(AMP)T7 T3 pBSR T3 T7ter pBudCE4.1(MCSI) CMV-Profor,T7 c-mycrev,EBVrevpBudCE4.1(MCSII) EF-1α Fwd BGHpBudCE4.1/lacZ/CAT CMV-Profor,T7 c-mycrev,EBVrev pBV220 pBV220F PBV220RpCAL-c T7/Tac-Profor T7-TerminatorpCAL-n T7/Tac-Profor T7-Terminator pCAMBIA1301(1300) pCAMBIA1301-5'(ECORI)/M13R pCAMBIA1301-3'(HindIII) pCAMBIA1304 pCAMBIA1301-5'(ECORI) pCAMBIA1301-3'(HindIII) pCAMBIA2300 M13R(-48) M13F(-47)/M13F(-49) pCANTB5E S1 S6pCAT3-enhancer RVP3 此载体⽆反向引物pCDFDuet-1(MCSI) ACYCDuetUP1 Primer DuetDOWN1pCDFDuet-1(MCSII) DuetUP2 T7TER pcDNA1.1 CMV-Profor,T7 pCDM8-3pcDNA2.1 M13for/T7 M13rev/Tac-ProforpcDNA3 T7/CMV-Profor BGH rev/SP6 pcDNA3.1(+/-) pcDNA3.1F/T7/CMV-Profor BGH rev pcDNA3.1/His(_A,_B,_C)T7/CMV-Profor BGH rev pcDNA3.1/myc-His(_A,_B,_C) T7/CMV-Profor BGH rev/c-mycrev pcDNA3.1/V5-His(_A,_B,_C) T7/CMV-Profor BGH rev/V5rev pcDNA3.1/Hygro(+,-) T7/CMV-Profor BGH rev/V5revpcDNA3.1/Zeo(+.-) T7 BGHrevpcDNA3.3TOPOTA CMV-Profor TKPAreverse pcDNA4/HisMax(_A,_B,_C) Xpressfor BGH revpcDNA4/HisMax-TOPO Xpressfor BGH rev pcDNA4/HisMax-TOPO/lacZ Xpressfor BGH rev pcDNA4/myc-His(_A,_B,_C) T7,CMV-Profor BGH rev,c-mycrev pcDNA4/TO TREfor/CMV-Profor BGH rev pcDNA4/V5-His(_A,_B,_C) CMV-Profor,T7 BGH rev,V5rev pcDNA4-TET/ON/AMP+ CMV-ProforpcDNA5/FRT T7/CMV-Profor BGH rev pcDNA6/myc-His(_A,_B,_C) CMV-Profor,T7 BGH rev,c-mycrev pcDNA6/V5-His(_A,_B,_C) CMV-Profor,T7 BGH rev,V5rev pCEP4 pCEP-F/CMV-Profor EBVrevpCI T7(17BASE)EBVrevpCI-neo T7(17BASE)EBVrev,T3pCMS-EGFP T7 T3pCMS-EGFP T7 EBVrev,T3pCMV-3Tag-4A T3 /PFLAG-CMV-F T7 pCMV5 pCMV5F PCMV5RpCMV6 CMV-Profor pCMV6-3、HGHrevpCMV6-XL4 CMV-Profor/T7 pCMV6-3、HGHrevpCMV-HA PCMV-F,CMV-Profor,TREfor PCMV-R,EBVrev pCMV-MYC(AMP+) PCMV-F PCMV-R, EBVrev pCMVmyc/nuc CMV-Profor BGH rev,c-mycrev pcmv-scriptEXVector T3 T7 pCMV-Sport6 T7/M13for SP6/M13rev pCOLADuet-1(MCSI) ACYCDuetUP1 Primer DuetDOWN1pCOLADuet-1(MCSII) DuetUP2 T7TER pCR2.1 T7,M13for M13rev pCR2.1-TOPO M13for,T7 M13rev pCR3.1 T7 BGH rev pCR3.1-Uni T7 BGH rev pCR4 T3/M13rev T7/M13for pCR4-Blunt T3/M13rev T7/M13forpCR4-TOPO M13rev,T3 M13for,T7pCR-Blunt M13rev T7,M13forpCR-BluntII M13rev/SP6 T7/M13for pCR-BluntII-TOPO M13rev/SP6 M13for/T7 pCRII M13rev/SP6 T7/M13for pCRII-TOPO M13rev/SP6 M13for/T7pCR-ScriptSK(+) T7/M13for T3/M13revpCR-XL-TOPO M13rev T7,M13forpCS2+(MCSI) SP6 EBVrev,T7?pCS2+(MCSII) T3 pSG5-3pDB_GAL4cam T7PpDB_leu PDBLeu-5 PDBLeu-3pDEST22 DEST22F DEST22RpDEST15 pGEX-5 T7-TerminatorpDEST20 pGEX-5 pFastBac-3pDEST26 TREfor/CMV-Profor T7/M13forpDisplay T7/CMV-Profor c-mycrev pDNR-LIB(MCS_A) M13for,T7 pSG5-3,M13R pDonar M13F M13RpDONR201 PDONR-F PDONR-RpDONR207 PDONR-F PDONR-RpDONR223 M13F M13RpDrive T7/M13rev SP6/M13for pDsRED1-C1 (DsRed1-N-f) DsRed1-C-rpDsRed1-N1 CMV-Profor RFP-Nrev,CMV-R?pDsRed2-N1 (DsRed1-N-f) DsRed1-C-rpDsRed2-C1 (DsRed1-N-f) DsRed1-C-r pDsRED2-C1(KAN+) pDsRED-ex-C1-F pEGFP-N-3’ pDsRed-ExpressdsRed1_N_primer/CMV-F dsRed1_C_primer/CMV-R pDSRED-monomer-N1(KAN+) dsRed1_N_primerpDSRED-N1(KAN+) pEGFP-N-5’ pDSRED-N-RpEASY-Blunt M13F T7/ M13RpEASY-BluntSimple M13F T7/ M13R(距离插⼊位点合适,优先安排)pEASY-T1 Simple M13F M13R(距离插⼊位点合适,优先安排)pEASY-T1 M13F/T7 M13RpEASY-T3 M13F/T7 M13R/SP6pECFP-N(C) PECFP-N-5. PECFP-N-3,pEF/myc/cyto/ER/mito/nuc pEF-5 Pcdna3.1R/BGH rev pEF1(4,6/myc-His) T7/pEF-5 Pcdna3.1R/BGH rev pEF1(4,6/V5-His) T7/pEF-5 Pcdna3.1R/BGH rev pEF1(4;6/His_A;_B;_C) T7/Xpressfor Pcdna3.1R/BGH rev pEF5/FRT/V5-D(-TOPO) T7/pEF-5 Pcdna3.1R/BGH rev pEGFP M13rev EGFP-NrevpEGFP-1 not available EGFP-Nrev/ PEGFP-N-3′pEGFP-C PEGFP-C-5′ PEGFP-C-3′pEGFP-C1 EGFP-Cfor SV40-pArevpEGFP-C2 EGFP-Cfor SV40-pArevpEGFP-C3 EGFP-Cfor SV40-pArevpEGFP-F EGFP-Cfor SV40-pArevpEGFP-N PEGFP-N-5′ PEGFP-N-3′pEGFP-N1 CMV-Profor EGFP-NrevpEGFP-N2 CMV-Profor EGFP-NrevpEGFP-N3 CMV-Profor EGFP-NrevpENTR M13F M13R pET-11(-a,-b,-c,-d) T7 T7-TerminatorpET-12(-a,-b,-c) T7 T7-Terminator pET-14b T7 T7-TerminatorpET-15b T7 T7-TerminatorpET-16b T7 T7-TerminatorpET-17b T7 T7-TerminatorpET-17xb T7 T7-TerminatorpET-19b T7 T7-TerminatorpET-20b(+) T7 T7-TerminatorpET-21(-a,-b,-c,-d)(+) T7 T7-Terminator pET-22b(+) T7 T7-TerminatorpET-23(-a,-b,-c,-d)(+) T7 T7-TerminatorpET-24(-a,-b,-c,-d)(+) T7 T7-Terminator pET-25b(+) T7 T7-TerminatorpET-26b(+) T7 T7-TerminatorpET-27b(+) T7 T7-Terminator pET-28a(-b,-c)(+) T7 T7-TerminatorpET-29a(-b,-c)(+) S.tag/T7 T7-TerminatorpET-3(-a,-b,-c,-d) T7 T7-Terminator pET-30_Ek/LIC T7 T7-Terminator pET-30_Xa/LIC T7 T7-Terminator pET-30a(-b,-c)(+) S.tag/T7 T7-Terminator pET-31b(+) not available T7-Terminator pET-32_Ek/LIC TRXfor T7-Terminator pET-32_XA/LIC TRXfor T7-Terminator pET-32a(-b,-c)(+) S.tag/TRXfor T7-Terminator pET-33b(+) T7 T7-Terminator pET-34b(+) S.tag T7-Terminator pET-35b(+) S.tag T7-Terminator pET-36b(+) S.tag T7-Terminator pET-37b(+) S.tag T7-Terminator pET-38b(+) not available T7-Terminator pET-39b(+) S.tag T7-Terminator pET-40b(+) S.tag T7-Terminator pET-41_Ek/LIC GST-Cfor T7-Terminator pET-41a(-b,-c)(+) S.tag/pGEX-5 T7-Terminator pET-42a(-b,-c)(+) S.tag/pGEX-5 T7-Terminator pET-43.1a(-b,-c) (+) S.tag coliDOWN pET-43.1a_EK/LIC S.tag not available pET-44a(-b,-c)(+) S.tag T7-Terminator pET-44a_EK/LIC S.tag not available pET-45b(+) T7 T7-Terminator pET-46_Ek/LIC T7 T7-Terminator pET-5(-a,-b,-c) T7 T7-Terminator pET-52b(+) T7 T7-Terminator pET-7 T7 T7-Terminator pET-9(-a,-b,-c,-d) T7 T7-Terminator pETcoco-1 T7 T7-Terminator pETDuet-1(MCSI) pETUP1 DuetDOWN1 pETDuet-1(MCSII) DuetUp2 T7-Terminator pET-T4 T7 T7-Terminator pEYFP-C1 pEGFP-C-5’pEGFP-C-3' pEYFP-N1 pEGFP-N-5’,CMV-Profor pEGFP-N-3’ pFLAG-ATS N26for pTrcHis-rv;C24rev pFLAG-CTC N26for pTrcHis-rv;C24rev pFLAG1 N26for pTrcHis-rv;C24rev pFLAG-CMV(-2) CMV-30;CMV-Profor CMV-24 pFLAG-CMV1 CMV-30 CMV-24pGAD424 GAL4_ADfor 3'ADpGADGH GAL4_ADfor 3'ADpGADT7 T7/GAL4_ADfor 3'AD pGADT7-Rec T7/GAL4_ADfor 3'AD pGADT-T T7 3,BD pGAD10 GAL4_ADfor 3'AD pGAPZa-A a-FACTOR 3'AOX pGBKT7 T7 3,BDpGBT9 MATCHMAKER5'DNA-BD/GAL4BD MATCHMAKER3'DNA-BDpGEM-1 T7 SP6pGEM-3 T7 SP6 pGEM-3Zf(+)(-) SP6/M13rev T7/M13for pGEM-4 T7 SP6pGEM-4Z T7/M13for SP6/M13rev pGEM-5Zf(+)(-) SP6/M13rev T7/M13for pGEM-7Zf(+)(-) SP6/M13rev T7/M13for pGEM-9Zf(+)(-) T7/M13for SP6/M13rev pGEM-T T7/M13for SP6/M13rev pGEM-T_Easy T7/M13for SP6/M13rev pGene/V5-His_A(_B,_C) pGENE-5 BGH rev,V5rev pGEX-2T pGEX-5 pGEX-3pGEX-2TK pGEX-5 pGEX-3pGEX-3X pGEX-5 pGEX-3 pGEX-4T-1(-2,-3) pGEX-5 pGEX-3 pGEX-5X-1(-2,-3) pGEX-5 pGEX-3 pGEX-6P-1(-2,-3) pGEX-5 pGEX-3 pGL2 GLP1 GLP2pGL3 RVP3 GLP2(RVP4)(没有特别说明⽤GLP2)pGL3-Basic(MCSI) RVP3(PGL3+)? GLP2(PGL3-)?pGL3-Basic(MCSII) RVP4, PLXSN-3'?pGL4pGlow(-TOPO) T7 pGlow-3 pGM-T T7 SP6pGWC SP6 T7pHook-2 T7/CMV-Profor not availablepIND/V5-His_A(_B,_C) not available BGH rev,V5rev PinpointTMVector PinpointPrimer SP6pIRES(MCSI) T7 pIRES-3pIRES(MCSII) not available T3,EBVrevpIRES2-EGFP CMV-Profor,pIRES2-EGFP.P5’ IRESrev,pIRES2-EGFP.P3’ pIRES-hrGFP-2a CMV-Profor,T3 pIRES-hrGFP-3 pIRESneo2 T7/CMV-Profor pIRES-3pIVEX(all) T7/pBRrevBam T7-TerminatorpLEXA pLEXA-F pLEXA-RpLNCX pLNCX-F pLNCX-RpLNCX2 CMV-Profor pRevTRE-3 pLVTHM(H1启动⼦下游)H1primer SP6 pLXSN pLXSN-F pLXSN-RpMALc MalE primer M13F(-47)pMAL-c2E MalE primer M13F(-47)pMAL-C2x Pmal-c2x-F(P5’) Pmal-C2X-R(P3’ )pMALd MalE primer M13F(-47)pMALe MalE primer M13F(-47)pMAL-P4X MalE primer M13F(-47)pMAL-p2X MalE primer M13F(-47)pMET A/B/C AUG1F AUG1RpMD18-T M13R(-48)ECORI M13F(-47)(HINDⅢ)pMD19-T M13F47 M13R48pMD20-T M13F47 M13R48pMOSBlue T7/M13rev M13forpMSCV-puro pLXSNfw pMSCVrev pMT/V5-His_A(_B,_C) Mtfor BGH rev,V5rev pNTAP T3 T7pCTAP pCTAP5'primer T7pOTB7 M13rev/T7 M13for/SP6 pOptiVEC-TOPO TA CMV-F/CMV-Profor EMCVIRES reverse pPC86 PPC86-F PPC86-R pPC97 PBD-GAL4-F PBD-GAL4-RpPIC3.5K 5'AOX 3'AOXpPic9k(A+,Z+) 5’AOX(Ppic9k-f)/a-factor 3’AOX(Ppic9k-R) pPICZa 5'AOX/PPICZa-F 3'AOX pPROEXHTA M13R(-48) ⽆反向引物pProEX-HTa(b,c) M13R48/ Tac-Profor pBADrv/pTrcHis-3 pPD129.36 M13F20/M13F pQE30or40or60(AMP+) PQE30-F PQE-30R pRC/CMV2/CAT CMV/T7pRECEIVER CMV-F 此载体⽆反向引物pRECEIVER-M03 pRECEIVER-M03F pRECEIVER-M03R pREP4 pREP4 forward primer EBV reverse primerpRK5 SP6 pSG5-3pRSET T7/Xpressfor T7ter pRSFDuet-1(MCSI) DuetUP1 DuetDOWN1 pRSFDuet-1(MCSII) DuetUP2 T7TER pSecTac2T7/CMV-Profor BGH rev/c-mycrevpSG5 pSG5-5,/T7 PSG5-R(SV40)pSHlox1 T7 SP6 pSHUTTLE-CMV PSHUTTLE-CMV-F PSHUTTLE-CMV-R pSI T7 EBVrev pSilencer1.0-U6 T7 T3pSilencer2.0-U6 T7 M13for pSilencer2.1-U6neo T7 M13for pSilencer3.1-H1hygro M13F(-47) 3.0REV pSilencer4_1-CMVneo pSilener4.1-5' CMV-Profor pSILENCEV2.0-U6 T7 2.0REV pSIneo T7 EBVrev,T3Psit T7 T7T pSK01-T M13F M13RpSK-CMV(KAN+) T7 T3 pSOS Psos5' Psos3'pSP64 SP6 M13revpSP64/65 SP6 此载体⽆反向引物pSP65 SP6 M13revpSP70 SP6 T7pSP71 SP6 T7pSP72 SP6 T7pSP73 SP6 T7pSport1 SP6/M13F(-47) T7/M13RpSPORT1 SP6/M13for T7/M13revpSPT18 SP6 T7pSTBlue T7/M13rev SP6/M13for pSUPER.Basic T7/M13F M13R pSUPER T7 T3pSURE-T M13F M13R pSURE-Tsimple M13F-47 M13R-48 pSuperCos pBRforEco T7pT3T7 M13rev/T3 M13for/T7pT7-7 T7 not availablepT7Blue? T7 M13F(-47)pT7T3_18U M13rev/T7 M13for/T3 pT7T3D-PAC M13rev/T7 M13for/T3 pTA2 M13F/T7(优先使⽤M13F)T3/M13RpT-Adv M13rev T7,M13forpTAg T7/M13rev M13for pTARGETTM pTarget.F(在T7前⾯)/T7 pTarget.R pTG19-T M13F(-47) M13R(-48) pTARGETTM T7/M13F PTARGET-SEQ pThioHisA,B,andC Trx Forward Trx Reverse / M13forpTO-T7 T7 T7TERpTRC99A PTRC99A-F pTrcHis-rvpTrc99a-c PTRC99C-F PTRC99C-RpTRC99a-c PTRC99C-F PTRC99C-RpTrcHisB PTrcHisA-F/Xpressfor PTrcHisA-RpTrcHisA PTrcHisA-F PTrcHisA-RpT-REX-DEST30 TREfor T7 pTriEx1 T7/TriExup TriExdownpTriEx1.1 T7/TriExup TriExdownpTriEx1.1_Hygro T7/TriExup not availablepTriEx1.1_Neo T7/TriExup IRESrevpTriplEX1 TriplEx-LD-5 T7,TriplEx-LD-3pTriplEX2 TriplEx-LD-5;pTR5‘ T7,TriplEx-LD-3pTWIN-1 T7 T7TERpTXB1 T7 MxeInteinrevpTXB2 T7 MxeInteinrevpTXB3 T7 MxeInteinrevpTYB1 T7 InteinrevpTYB2 T7 InteinrevpTYB3 T7 InteinrevpTYB4 T7 InteinrevpTZ_18U T7/M13rev M13forpUB6/V5-His UB-F BGH rev,V5rev pUC13 M13rev/M13rev(-48) M13for/M13for(-49)pUC18 M13rev/M13rev(-48) M13for/M13for(-49) pUC18(19)/118(119) M13F(-47) M13R(-48)/PUC19(HINDⅢ) pUC19M13rev/M13rev(-48) M13for/M13for(-49)pUC57 M13rev/M13rev(-48) M13for/M13for(-49)pUC8 M13rev/M13rev(-48) M13for/M13for(-49)pUC9 M13rev/M13rev(-48) M13for/M13for(-49)pUM-T M13F(-20) M13R(-20)pUCm-T(AMP+) M13F/T7 M13RpUNI/V5-His not available BGH rev,V5revpVAX1T7 T7/CMV-Profor PCDNA3.1R/BGHR pVP22/mycHis VP22-Cfor BGH rev,c-mycrev pVP22/mycHis2 T7/CMV-Profor VP22-Nrev pWE15 pBRrevHind T7pYES2 T7 pYes2.R pYES-DEST52 T7/GAL1-Profor V5rev/CYC1-Terminator pYEX_4T_1 Clontech pYEX_4T-3pYEX_4T_2 pGEX-5 pYEX_4T-4pYEX_4T_3 pGEX-5 pYEX_4T-4pYEX_4T_4 pGEX-5 pYEX_4T-4 pZeoSV2+/hu-Tyr T7 not available pZERO-1 M13rev/SP6 T7/M13forpZERO-2 M13rev/SP6 M13for/T7pZome-1-C Zome-F Zome-RpZome-1-N Zome-F Zome-R YCp50(BamHI-site) PBRforBam PBRrevBam YEp13(BamHI-site) PBRforBam PBRrevBam YEp24(BamHI-site) PBRforBam PBRrevBam YIp5(BamHI-site) PBRforBam PBRrevBam RNAi-ReadypSIREN-RetroQ-U6ZsGreen U6菌PCDB T7 BGH 天为时代TA载体T7 T3 m13(+)T7?T3?pMSCV-neo pMSCV-5' pMSCV-3'。
引物相关知识点总结大全

引物相关知识点总结大全一、引物的概念引物是指用于核酸杂交、PCR、定量PCR、DNA测序以及RNA分子的反向转录(RT)等实验技术中的一种短链DNA或RNA,通常是15-30个核苷酸的长度。
引物的主要作用是与目标序列中的互补序列形成双链结构,从而引发特定的生物学反应。
在实验室中,引物通常由合成核酸技术制备而成。
二、引物的分类根据引物作用的目标序列不同,引物可分为DNA引物和RNA引物两种。
DNA引物主要用于PCR、序列特异性分析等实验中,而RNA引物则主要用于RT-PCR等实验技术中。
根据引物的长度和作用位置不同,引物可以分为引导引物和扩增引物。
引导引物通常用于核酸杂交、PCR等实验中,用于可选择的结合目标序列,一般长度为15-30个碱基。
扩增引物则主要用于PCR实验中,其长度和序列将直接影响到PCR扩增的结果。
三、引物的设计原则1. 引物的互补性:DNA引物或RNA引物与待扩增的目标序列的互补性是引物设计的基本原则,引物与目标序列要求完全互补,并且在同一温度下具有相似的Tm值。
2. 避免引物二聚体和含量:引物的设计应避免引物自身的二聚体和寡聚体的产生,并且要避免引物的富含量,以提高PCR扩增的效率和特异性。
3. 引物的特异性:引物的设计要求尽量确保引物特异性,即引物只与目标序列特异性结合,不与其他非靶标序列结合。
4. 引物长度和GC含量:引物的长度和GC含量对PCR扩增的特异性和效率有一定的影响,一般长度在18-25个碱基,GC含量在40-60%之间。
5. 引物的5’端末修饰:引物的5’端末修饰,如磷酸化和生物素化等,可以增强引物与酶和材料的连接性和稳定性。
四、引物的应用引物作为实验室技术中的重要工具,在许多生物学实验中都扮演着重要的角色。
其主要应用包括:1. PCR扩增:PCR扩增是引物最常见的应用之一,引物特异性与目标序列的互补性可以实现PCR扩增特定基因或序列。
2. DNA测序:引物也常用于DNA测序技术中,引物和DNA降解酶一起作用可以在酶刻蚀DNA链的末端,并且通过引物的特异性与酶反应的DNA片段在随后的扩增和测序中得到全面和特异性的测序结果。
菌种鉴定通用引物

菌种鉴定通用引物引言菌种鉴定是微生物学领域中的一项重要工作,它对于疾病诊断、环境监测和食品安全等方面具有重要意义。
菌种鉴定的关键是通过引物对目标微生物的DNA序列进行扩增和分析,从而确定其分类地位和种属鉴定。
本文将介绍菌种鉴定通用引物的原理和应用。
一、引物的选择原则菌种鉴定通用引物的选择需要考虑以下几个因素:1. 引物的特异性:引物应具有足够的特异性,只能扩增目标微生物的DNA序列,而不扩增其他非目标微生物的DNA序列。
2. 引物的保守性:引物应选择在目标微生物的DNA序列中高度保守的区域,以确保扩增的目标序列的一致性。
3. 引物的长度:引物的长度应适中,一般为18-30个碱基对,过长的引物可能导致扩增效率降低,而过短的引物可能导致特异性降低。
4. 引物的G+C含量:引物的G+C含量应适中,一般在40-60%之间,过低或过高的G+C含量可能影响扩增效率。
5. 引物的互补性:引物的两个端部应互补,以确保引物在目标DNA 序列上的结合和扩增。
二、常用的菌种鉴定通用引物1. 16S rRNA通用引物:16S rRNA序列是细菌和古菌中高度保守的基因序列,因此广泛用于细菌和古菌的鉴定。
常用的引物包括27F(5'-AGAGTTTGATCCTGGCTCAG-3')和1492R(5'-GGTTACCTTGTTACGACTT-3')。
2. 18S rRNA通用引物:18S rRNA序列是真核生物中高度保守的基因序列,因此广泛用于真菌和原生动物的鉴定。
常用的引物包括ITS1(5'-TCCGTAGGTGAACCTGCGG-3')和ITS4(5'-TCCTCCGCTTATTGATATGC-3')。
3. 基因编码区通用引物:除了rRNA序列外,一些基因编码区的引物也被广泛应用于菌种鉴定。
例如,ITS区(内转录间隔区)是真菌常用的鉴定区域,常用的引物包括ITS1和ITS4。
常用载体及相关通用引物

常用载体及相关通用引物默认分类2008-10-06 13:16:31 阅读512 评论0 字号:大中小载体一端另一端pBlueScriptSK(+) M13F(-77)/M13F(-47)/M13F/T7 M13R(-88)/M13R(-48)/M13R/T3 pcDNA3 H1.3F/pCMV-F /pEGFP-N-5/T7 BGHpcDNA3.1(+) H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.0(+)/myc-His C/A/B H1.3F/pCMV-F /pEGFP-N-5/T7 BGHpcDNA3.1(+)/myc-His C/A/B H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1(+)/myc-His/LacZ H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/His C/A/B H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/His/LacZ H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/Hygro(-) H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/Hygro(+) H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/Hygro/LacZ H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/V5-His C/A/B H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/V5-His/LacZ H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/V5-His-TOPO H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/V5-His-TOPO/LacZ H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/Zeo(-) H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/Zeo(+) H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpcDNA3.1/Zeo/CAT H1.3F/pCMV-F /pEGFP-N-5/T7 pCDNA3.1R/BGHpColdI DNA p-GEX3’pColdII DNA p-GEX3’pColdIII DNA p-GEX3’pColdIV DNA p-GEX3’pDream2.1 H1.3F/pCMV-F /pEGFP-N-5/T7 SP6pEGFP-N1,2,3 H1.3F/pCMV-F /pEGFP-N-5 pEGFP-N-3’/pEGFP-C-5’/pEGFP-C-3' pET15b T7 T7terpET20b T7 T7terpET22b T7 T7terpET28a T7 T7terpET28b T7 T7terpET30b T7 T7terpFastBac Dual pEGFP-C-3'pFastBac HT A,B,C pEGFP-C-3'pFastBac1 pEGFP-C-3'pGEM-T Easy M13F(-77)/M13F(-47)/M13F/T7 M13R(-88)/M13R(-48)/M13RpGEX-4T-1/3 p-GEX5’ p-GEX3’pGS-21a p-GEX5’ T7terpLenti6/V5-D-TOPO M13F(-47)/M13F/T7 M13R(-48)/M13R/T3pLenti6/V5-GW/lacZ M13F(-47)/M13F/T7 M13R(-48)/M13R/T3pMAL-p2x M13F(-47)/M13F/PBV220R M13R(-48)pMD18-T M13F(-77)/M13F(-47)/M13F M13R(-88)/M13R(-48)/M13RpMD19-T M13F(-77)/M13F(-47)/M13F M13R(-88)/M13R(-48)/M13RpMTBiPV5-His GFP p-GEX3’/ M13F(-47)/M13F M13R(-88)/M13R(-48)/M13R /pEGFP-C-3'/BGH pPCR script M13F(-77)/M13F(-47)/M13F/T7 M13R(-88)/M13R(-48)/M13R/T3pPIC3.5 5’AOX 3’AOXpPIC9 5’AOX 3’AOXpPICZ C/A/B 5’AOX 3’AOXpPICZalpha C/A/B 5’AOX/α-Factor 3’AOXpSecTag2 A,B,C H1.3F/pCMV-F /pEGFP-N-5/T7 BGHpTWIN T7 T7terpUC57 M13F(-77)/M13F(-47)/M13F M13R(-88)/M13R(-48)/M13RpUC18 M13F(-47)/M13F M13R(-48)/M13RpUC19 M13F(-47)/M13F M13R(-48)/M13R其他…… ……注:1)排列顺序为:有左到右==(离插入为点)由远到近;2)若有多个引物,红色的为首选引物:3)选择测序通用引物时,以距离插入位点约50碱基为佳;4)同时有T7,T7ter的载体,单反应首选T7ter;5)同时有T7,pCDNA3.1R的载体,单反应首选pCDNA3.1R ;6)同时有T7,BGH的载体,单反应首选BGH;7)同时有pCMV-F,BGH的载体,单反应首选BGH;。
常用载体测序引物列表

常用载体测序引物列表 载体名称 正向引物 反向引物pACT T7 T3pACT2 GAL4 AD(5’AD) 3'AD/pACT2-R PACYCDUET-1(MCSII) DuetUP2 T7TERpAD/pl-DEST CMV-FpAD-Track pAD-Track-F 无pAS2-1 5’BD M13-26/48 PB42AD PB42ADF PB42ADR PBAD PBAD-F PBAD-R pBABE pBABE5’ pBABE3’ PBACPAK8 BAC1BAC2 pBK-CMV T7/M13-20 T3/M13-26 PBS(SK/KS)/M13- M13F/T7T3/M13R pBV220 PBV220F PBV220R pBI121需根据酶切位点确认 需根据酶切位点确认 pCAMBIA 1301(1300)需根据酶切位点确认 需根据酶切位点确认 pCAMBIA 2300 需根据酶切位点确认 需根据酶切位点确认PCANTB5E S1/M13R S6pCAT3-enhancer pGL3+pcDNA3.0 pEGFP-N5/T7 SP6/BGHpcDNA3.1 pEGFP-N5/T7 BGHpcDNA4 T7/pEGFP-N5 BGHpcDNA6 T7/pEGFP-N5 BGHpcDNAII T7SP6 pCE2.1 M13FM13R pCEP4 pCMV5R/PCEP-F pEGFP-N5/EBV-R pCF-T M13FM13R pCIT7(17Base ) pCMV5R pCI-neo T7(17Base ) T3pCMS-EGFP T7T3 pCMV-3Tag-4A T3 /pFlag-CMV-F T7pCMV5 pCMV5F pCMV5R pCMV5-Flag pCMV5FpCMV5R pCMV-MYC/NUC pCMV5FpCMV5R PCMV-HA pCMV5FpCMV5R pCMV-SportSP6/M13RpCMV-Tag T3T7 pCMV-TNT T7/SP6pCMV5R pCR2.1-TOPO M13F/T7M13R pCR3.1 T7BGH pCS2 SP6T7PDNR-LIB M13F M13RpDonar M13F M13R pDONR201 pDONR-F pDONR-R pDONR221 M13F M13RpDrive T7 SP6 pDRIveR T7 SP6pDsRED1-C1 pDsRED-C1-F pECFP-C-3 pDsRed1-N1 pEGFP-N-5 RFP-NrevpDsRed-Monomer-N1 pEGFP-N-5 pDsRed1-NpDsRED2-C1 pDsRED-ex-C1-F pEGFP-N-3’pEGFP-N-5 pDsRed1-N pDSRED2-N1pDSRED-N1 pEGFP-N-5’ PDSRED-N-R pEASY-BLUNT M13F/T7 M13RTerpEASY-E1 T7 T7pEASY-T1 M13F/T7 M13RpEASY-T3 M13F/T7 M13R/SP6 pECFP-C pEGFP-C-5’ pECFP-C-3' pECFP-N1 pEGFP-N-5’ pEGFP-N-3’pEF/myc/ER pEF-F pCDNA3.1R pEGFP-C pEGFP-C-5’ pEGFP-C-3' pEGFP-N pEGFP-N-5’ pEGFP-N-3’ pENTR/D-TOPO M13F M13RTerpET-*(His) T7 T7pET22(a,b,c) T7 T7TerTerpET28,30,24,49 T7 T7TerpET32(a,b,c) T7/S.tag T7pET-42a (-b, -c)(+) Stag T7 TerpET50 T7/s.tag T7TERpET44 T7/s.tag T7Ter pEYFP-N1 pEGFP-N-5’ pEGFP-N-3’ PETBlue M13-20 PETBlueDOWN PETDUET-1(MCSI) PBRrevBam DuetDOMN1PETDUET-1(MCSII) DuetUP2 T7TERpEYFP-C1 pEGFP-C-5’ pEGFP-C-3' pEYFP-N1 pEGFP-N-5’ pEGFP-N-3’ pFastBac pFastBac-F pFastBac-R pFastBac HT_A,B,C pFastBac-PH pECFP-C-3 pFastBac1 pFastBac-PH pECFP-C-3 pFastBac Dual (MCSI) pFastBac-PH pECFP-C-3 pFLAG-CMV CMV30/pFLAG-CMV-F CMV-24/pFLAG-CMV-Rp3xFlag-CMV-7.1 CMV-F/CMV24 CMV30pGAD424 5’AD 3'ADpGADGH 5’AD 3'ADpGADT7-Rec 5’AD/T7 3'ADpGAPZa-A/B/C a-FACTOR 3'AOXforward 3'AOXpGAPZ-A/B/C pGAPpGBKT7 T7 3'BDpGEM-T(-Easy) M13F/T7 SP6/M13RpGEX-(*)T pGEX-5’ pGEX-3’pGL2 pCMV5R PGL3-Pgl3-BASIC pGL3+ PGL3-pGL3-Enhancer pGL3+ PGL3-PinPoint TM PinPoint Primer SP6pIRES PIRES-F/T7 T3/PIRES-RpIRES2-EGFP pIRES2-EGFP.P5’ pIRES2-EGFP.P3’ pLenti6/v5-Dest pEGFP-N-5’PLEXA PLEXA-F PLEXA-R PLNCX PLNCX-F PLNCX-R pLXSN pLXSN-F pLXSN-RPjc1-tac T7pJet.1.21.Blunt T7primer M13F(-47) pMAL-c2E MalEP3/PMAL-C2X-R pMAL-C2x P5/PMAL-C2X-Fprimer M13F(-47) pMAL-p2X MalEPMD18-T M13R(-48) M13F(-47) PMD19-T M13F(-47) M13R(-48) pMIR-report M13FPPC86 PPC86-F PPC86-R3’AOXpPIC9K 5’AOX/a-factor3'AOX pPICZa 5'AOX/PPICZa-FpPROEXHTA M13R(-48)pQE30or40 pQE30+ pQE30- pRECEIVER CMV-FpRSET T7 T7TER PRSFDUET-1(MCSI) DuetUP1 DuetDOWN1 PRSFDUET-1(MCSII) DuetUP2 T7TERpShuttle-CMV pShuttle-CMV-F pShuttle-CMV-R PSILENCE-1.0-U6 T7 T3pSilencer3.1-H1 hygro M13F(-47) 3.0revpSILENCEV2.0-U6 T7 2.0revpSK01-T M13F M13RpSK-CMV T7 T3Psos H1PSP64/65 SP6pSP72 T7 SP6T7/M13RpSport1 SP6/M13F(-47)pSTBlue-1 T7 SP6pSUPER T7 T3pT7Blue(R) T7 M13F(-47)pTA2 M13F/T7 T3/M13R/T7 M13RpT-Adv M13FpTARGET TM pTarget-F/T7 arget-RPTHIOHISA,B,C TRX-FORWARD TRX-REVERSEpTO-T7 T7 T7TERPBV220-/PTRC99C-R pTRC99a-c PQE30+/PTRC99C-FpTriPLEx2 PT5/5'TriplEx2 T7pTz57r/t M13+ M13-pTWIN-1 T7 T7TERpUC18(19)/118(119) M13F(-47) M13R(-48)M13F(-47)/M13F pUC57 M13R(-48)/M13RpUCm-T M13F/T7 M13RpVAX1 T7 BGHT3/M13R/M13R(-48) pWSK29 T7/M13F/M13F(-47)pXT7 T7 SP6pYES2 T7 pYes2.R。
测序通用引物序列

测序通用引物序列常见载体的测序引物:Primer of Vector:Vector:Primer(F);Primer(R)pACT T7 T3pACT2 GAL4 AD pACT2-RpAS2-1 GAL4 BD pAS2-1.RpB42AD pB42ADF pB42ADRpBACPAK8 BAC1 BAC2pBK-CMV T7 T3PBS(SK/KS)/M13- M13F/T7 T3/M13RpBV220 PBV220F PBV220RpCAMBIA 1301(1300) P1 P2pCAMBIA 2300 M13R(-48) M13F(-47)PCANTB5E S1/M13R S6pCAT3-enhancer RVP3 此载体无反向引物pcDNA3.0 CMV-F/T7 SP6/BGHpcDNA3.1 T7 BGHpcDNA4 T7/CMV-F BGHpcDNA6 T7/CMV-F(J21025在T7前面)BGHpcDNAII T7 SP6pCE2.1 M13F M13RpCEP4 pCEP-F EBV-RpCF-T M13F M13RpCI T7(17Base)此载体无反向引物pCI-neo T7(17Base)T3pCMS-EGFP T7 T3pCMV-3Tag-4A T3 /PFLAG-CMV-F T7pCMV5 pCMV5F pCMV5RpCMV5-Flag pCMV5F pCMV5RpCMV-MYC/HA pCMV-F pCMV-RpCMV-Sport M13F/T7 SP6/M13RpCMV-Tag(KAN+) T3 T7pCR2.1-TOPO M13F/T7 M13RpCR3.1 T7 BGHpCS2 SP6 T7pDonar M13F M13RpDONR221 M13F M13RpDrive T7 SP6pDRIveR(KAN+) T7 SP6pDsRED1-C1 pDsRED-ex-C1-F pEGFP-N-3’(距离很近,一般不用) pDsRED2-C1(KAN+) pDsRED-ex-C1-F pEGFP-N-3’pDSRED-N1(KAN+) pEGFP-N-5’ PDSRED-N-RpECFP-C pEGFP-C-5’ pECFP-C-3′pECFP-N1 pEGFP-N-5’ pEGFP-N-3’pEF/myc/ER pEF-F pCDNA3.1RpEGFP-C(KAN+) pEGFP-C-5’ pEGFP-C-3′pEGFP-N(KAN+) pEGFP-N-5’ pEGFP-N-3’pENTR/D-TOPO(KAN+) M13F M13RpET-*(His)(KAN+) T7 T7 TerpET22(a,b,c)(AMP+) T7 T7 TerpET28,30,24,49(KAN+) T7 T7 TerpET32(a,b,c)(AMP+) T7/S.tag T7 TerpET50(amp+) T7/s.tag T7TERpET44(AMP+) T7/s.tag T7 TerpEYFP-N1 pEGFP-N-5’ pEGFP-N-3’pEYFP-C1 pEGFP-C-5’ pEGFP-C-3′pFLAG-CMV pFLAG-CMV -F pFLAG-CMV -R pGAD424 GAL4 AD 3′ADpGADGH GAL4 AD 3′ADpGADT7-Rec GAL4 AD/T7 3′ADpGAPZa-A a-FACTOR 3′AOXpGBKT7(Kana+) T7 3′BDpGEM-T(-Easy) M13F/T7 SP6/M13RpGEX-(*)T pGEX-4T-5’ pGEX-4T-3’pGL2 GLP1 GLP2pGL3 RVP3 GLP2(RVP4)(没有特别说明用GLP2) PinpointTM Vector Pinpoint Primer SP6pIRES PIRES-F/T7 T3/PIRES-R (J40720)pIRES2-EGFP pIRES2-EGFP.P5’ pIRES2-EGFP.P3’pLXSN pLXSN-F pLXSN-RpMAL-c2E MalE primer M13F(-47)pMAL-C2x P5’ P3’pMAL-p2X MalE primer M13F(-47)PMD18-T M13R(-48) M13F(-47)PMD19-T M13F(-47) M13R(-48)pPIC9K(AMP+、ZEO+)5’AOX/a-factor 3’AOX pPICZa 5′AOX/PPICZa-F 3′AOXpPROEXHTA M13R(-48) 无反向引物pQE30or40 pQE30-F pQE-R:pRSET T7 T7TERpSilencer3.1-H1 hygro M13F(-47) 3.0REV pSILENCEV2.0-U6 T7 2.0REVpSK01-T M13F M13RpSK-CMV(KAN+) T7 T3pSP72 T7 SP6pSport1 SP6/M13F(-47) T7/M13RpSUPER T7 T3pTA2 M13F/T7(优先使用M13F)T3/M13RpT-Adv M13F /T7 M13RpTARGET TM pTarget.F(在T7前面)/T7 pTarget.R pTO-T7 T7 T7TERpTRC99a-c pTRC99C-F pTRC99C-RpTriPLEx2 5′pTriPLEx2 T7pTWIN-1 T7 T7TER(客户确认再用)pUC18(19)/118(119) M13F(-47) M13R(-48)pUCm-T M13F/T7 M13RpVAX1 T7 BGHpWSK29 T7/M13F/M13F(-47) T3/M13R/M13R(-48)pXT7 T7 SP6pYES2 T7 pYes2.RpLEXA pLEXA-F pLEXA-RpLNCX pLNCX-F pLNCX-RpRECEIVER CMV-F 此载体无反向引物pSHUTTLE-CMV PSHUTTLE-CMV-F PSHUTTLE-CMV-R pSP64/65 SP6 此载体无反向引物pSTBlue-1 T7 SP6pT7Blue(R) T7 M13F(-47)引物名称序列(5′-3′):M13R:CAG GAA ACA GCT ATG ACCM13F:TGT AAA ACG ACG GCC AGTM13F(-47):CGC CAG GGT TTT CCC AGT CAC GACM13R(-48):AGC GGA TAA CAA TTT CAC ACA GGAM13(-96):CCC TCA TAG TTA GCG TAA CGSP6:ATT TAG GTG ACA CTA TAGT7:TAA TAC GAC TCA CTA TAG GGT7 terminator:TGC TAG TTA TTG CTC AGC GGT3:ATT AAC CCT CAC TAA AGG GApGEX-4T-5′:GGG CTG GCA AGC CAC GTT TGG TG pGEX-4T-3′:CCG GGA GCT GCA TGT GTC AGA GG GLp1:TGT ATC TTA TGG TAC TGT AAC TGGLp2:CTT TAT GTT TTT GGC GTC TTC CARVp3:CTA GCA AAA TAG GCT GTC CCRVp4:GAC GAT AGT CAT GCC CCG CGpcDNA3.1R:TAG AAG GCA CAG TCG AGGPinPoint primer:CGT GAC GCG GTG CAG GGC G pCMV-F:TCT AAA AGC TGC GGA ATT GTpCMV-R:TCCAAACTCATCAATGTATCpTRC99C-F: TTG CGC CGA CAT CAT AACpTRC99C-R: CTGCGTTCTGATTTAATCTGpCEP-F: AGA GCT CGT TTA GTG AAC CGEBV-R : GTG GTT TGT CCA AAC TCA TCpIRES2-EGFP.P5’: GTA GGC GTG TAC GGT GGG AG pIRES2-EGFP.P3’: AAC GCA CAC CGG CCT TAT TC3′AD: AGA TGG TGC ACG ATG CAC AGCMV -F:CGC AAA TGG GCG GTA GGC GTGS1:CAA CGT GAA AAA ATT ATT ATT CGCS6:GTA AAT GAA TTT TCT GTA GTA GG5`AOX1:GAC TGG TTC CAA TTG ACA AGC3`AOX1:GCA AAT GGC ATT CTG ACA TCCα-Factor:TAC TAT TGC CAG CAT TGC TGCGAL4 AD:TAC CAC TAC AAT GGA TGpACT2-R:GTGCACGATGCACAGTTGAApB42ADF:CCA GCC TCT TGC TGA GTG GAG ATGpB42ADR:AAG CCG ACA ACC TTG ATT GGA GpEGFP-N-5’:TGG GAG GTC TAT ATA AGC AGA G pEGFP-N-3’:CGT CGC CGT CCA GCT CGA CCA G pEGFP-C-5’:CAT GGT CCT GCT GGA GTT CGT G pEGFP-C-3′ :TAT GGC TGA TTA TGA TCA GTPBV220F:AAG AAG GGC AGC ATT CAA AGPBV220R:CTG CGT TCT GAT TTA ATC TGU6:ATG GAC TAT CAT ATG CTT ACC GTA2.0rev primer:AGG CGA TTA AGT TGG GTA3.0rev:GAG TTA GCT CAC TCA TTA GGCS.tag:GAA CGC CAG CAC ATG GAC5′TriplEx2:CTC CGA GAT CTG GAC GAG CRVP4:GAC GAT AGT CAT GCC CCG CGRVP3:CTA GCA AAA TAG GCT GTC CCpQE30-R:GTT CTG AGG TCA TTA CTG GpQE30-F:TGA GCG GAT AAC AAT TTC ACpEF-F:TCA AGC CTC AGA CAG TGG TTCpDSRED-N1-R:TGA AGC GCA TGA ACT CCT TG pDsRED-express-C1-F:TCC CAC AAC GAG GAC TAC AC pCMV5R:ATT ATA GAG GAC ACC TAG TCpCMV5F:TTC CAA AAT GTC GTA ATA ACP5′:TGC GTA CTG CGG TGA TCA ACP3′:CTG CAA GGC GAT TAA GTT GGBAC2:ACG CAC AGA ATC TAG CGC TTBAC1:AAC CAT CTC GCA AAT AAA TA3′BD:TAA GAG TCA CTT TAA AAT TTG TAT AC pTarget.F:CCA GGA TTT TCC CAG TCA CpTarget.R:GGC TTT ACA CTT TAT GCT TCT7(17base):ACA TCC ACT TTG CCT TTC TCpYes2.R:TCG GTT AGA GCG GAT GTGGAL4 BD:TCA TCG GAA GAG AGT AGpAS2-1.R:AAG AGT TAC TCA AGA ACA AGA ApFlag-CMV-F: GGT AGG CGT GTA CGG TGGpFlag-CMV-R: GCA CTG GAG TGG CAA CTTpIRES-F: CTT ACT GAC ATC CAC TTT GCpIRES.R: CAC TGC ATT CTA GTT GTG GTP1:CCA GGC TTT ACA CTT TAT GCP2:GCG ATT AAG TTG GGT AAC GCpLEXA-F:CGT CAG CAG AGC TTC ACC ATpLEXA-R:TAA AAC CTA AGA GTC ACT TTpLNCX-F:AGC TCG TTT AGT GAA CCG TCA GAT CG pLNCX-R:ACC TAC AGG TGG GGT CTT TCA TTC CC pLXSN-F:CTT GAA CCT CCT CGT TCG ACpLXSN-R:GTT GCT GAC TAA TTG AGA TGpSHUTTLE-CMV-F:GGT CTA TAT AAG CAG AGG TG pSHUTTLE-CMV-R:GTG GTA TGG CTG ATT ATG ATC AG MalE Primer:GGT CGT CAG ACT GTC GAT GAA GCC。
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3’AOX5′-GGCAAATGGCATTCTGACAT-3‘3‘AD5′- AGA TGG TGC ACG ATG CAC AG-3‘3‘BD5′-TAA GAG TCA CTT TAA AAT TTG TAT C-3‘5’AOX5′-GACTGGTTCCAATTGACAAGC-3‘A-FACTOR5′-TAC TAT TGC CAG CAT TGC TGC-3‘CMV-F5′-CGCAAATGGGCGGTAGGCGTG-3‘CMV-R5′-GTTCACGGTGCCCTCC-3‘EGFP-Nrev5′-CGT CGC CGT CCA GCT C-3‘GLP15′-TGT ATC TTA TGG TAC TGT AAC TG-3‘GLP25′-CTT TAT GTT TTT GGC GTC TTC CA-3‘M13(-96)5′-CCCTCATAGTTAGCGTAACG-3‘M13-205′-GTA AAA CGA CGG CCA GTG-3‘M13F5′-TGTAAAACGACGGCCAGT-3‘M13F(-47)5′-CGCCAGGGTTTTCCCAGTCACGAC-3‘M13F-215′-TGT AAA ACG ACG GCC AGT-3‘M13R5′-CAGGAAACAGCTATGACC-3‘M13R(-48)5′-AGCGGATAACAATTTCACACAGGA-3‘P125845′-TTTTCAGTATCTACGAT-3’P171105′-TACCACTACAATGGATG-3’pbd-gal4-f5′-GCCTCTAACATTGAGACAGC-3‘pbd-gal4-r5′-AAGAGTTACTCAAGAACAAGAA-3‘pbi-121(35s)5′-GACGCACAATCCCACTATCC-3‘PBV220F5′-AAGAAGGGCAGCATTCAAAG-3‘PBV220R5′-CTG CGT TCT GAT TTA ATC TG-3‘PCDNA3.0-R25′-GGC AAC TAG AAG GCA CAG TC-3‘PCDNA3.1F5′-CTAGAGAACCCACTGCTTAC-3’PCDNA3.1R5′-TAGAAGGCACAGTCGAGG-3’PCMV-F5′-TCTAAAAGCTGCGGAATTGT-3‘PCMV-5F5′-TTCCAAAATGTCGTAATAAC-3‘PCMV-R5′-TCCAAACTCATCAATGTATC-3‘PCMV-5R5′-ATTATAGAGGACACCTAGTC-3‘PEGFP-C-3’5′-TATGGCTGATTATGATCAGT-3‘PEGFP-C-5’5′-CATGGTCCTGCTGGAGTTCGTG-3‘PEGFP-N-3’5′-CGTCGCCGTCCAGCTCGACCAG-3‘PEGFP-N-5’5′-TGGGAGGTCTATATAAGCAGAG-3‘PGEX-3’5′-CCGGGAGCTGCATGTGTCAGAGG-3‘PGEX-5’5′-GGGCTGGCAAGCCACGTTTGGTG-3‘PIRES2-EGFP-F5′-GTA GGC GTG TAC GGT GGG AG-3‘PIRES3-EGFP-R5′-AAC GCA CAC CGG CCT TAT TC-3‘pMAL-C2X-F5′-TGC GTA CTG CGG TGA TCA AC-3’pMAL-C2X-R5′-CTG CAA GGC GAT TAA GTT GG-3’PQE30F5′- TGA GCG GAT AAC AAT TTC AC-3‘PQE30R5′- GTT CTG AGG TCA TTA CTG G-3‘RVP35′-CTA GCA AAA TAG GCT GTC CC-3‘RVP45′-GAC GAT AGT CAT GCC CCG CG-3‘S.TAG5-CGAACGCCAGCACATGGACA-3‘SP65-ATTTAGGTGACACTATAG-3‘T35-ATTAACCCTCACTAAAGGGA-3‘T75‘-TAATACGACTCACTATAGGG-3‘T7(17BASE)5‘-ACATCCACTTTGCCTTTCTC-3‘T7TER5‘-TGCTAGTTATTGCTCAGCGG-3‘NOSR AAT CAT CGC AAG ACC GGCM13 Forward (-20) 5'-GTA AAA CGA CGG CCA GTG-3'M13 Reverse (-27) 5'-GGA AAC AGC TAT GAC CAT G-3'M13 Forward (-47) 5'-CGCCAGGGTTTTCCCAGTCACGAC-3'M13 Reverse (-48) 5'-AGC GGA TAA CAA TTTC ACA C-3'SP6 5'-TAC GAT TTA GGT GAC ACT ATA G-3'T3 5'-CAA TTA ACC CTC ACT AAA GG-3'T7 5'-TAA TAC GAC TCA CTA TAG GG-3'T7Term 5'-GCT AGT TAT TGC TCA GCG G-3' pBluescript KS 5'-TCG AGG TCG ACG GTA TC-3' pBluescript SK 5'-CGC TCT AGA ACT AGT GGA TC-3'3'pGEX 5'-CCG GGA GCT GCA TGT GTC AGA GG-3' 5'pGEX 5'-GGG CTG GCA AGC CAC GTT TGG TG-3' GST-Tag 5'-ACC CAA TGT GCC TGG ATG CG-3' pTrcHis-Forward 5'-GAG GTA TAT ATT AAT GTA TCG-3' pTrcHis-Reverse 5'-GAT TTA ATC TGT ATC AGGCMV-Forward 5'-CGC AAA TGG GCG GTA GGC GTG-3' CMV-Reverse 5'-AGT AGG AAA GTC CCG TAA GG-3' EGFP-C 5'-CAT GGT CCT GCT GGA GTT CGT G-3' EGFP-N 5'-CGT CGC CGT CCA GCT CGA CCA-3' BGH-Reverse 5'-TAG AAG GCA CAG TCG AGG-3' pQEproseq 5'-CCC GAA AAG TGC CAC CTG-3' pQErevseq 5'-GTT CTG AGG TCA TTA CTG G-3'Intein Forward 5'-CCC GCC GCT GCT TTT GCA CGT GAG-3' 5'-pBabe-Seq 5'-CTT TAT CCA GCC CTC AC-3'3'-pBabe-Seq 5'-ACC CTA ACT GAC ACA CATT CC-3'-96 glll Sequencing Primer 5'-CCC TCA TAG TTA GCG TAA CG-3'GAL1 Forward 5'-AAT ATA CCT CTA TAC TTT AAC GTC-3' pBAD Forward 5'-ATG CCA TAG CAT TTT TAT CC-3'pBAD Reverse 5'-GATTTAATCTGTATCAGGpTRE 3' 5'-CCA CAC CTC CCC CTG AAC-3'pTRE 5' 5'-CGC CTG GAG ACG CCA TCC-3' pYESTrp Forward 5'-GAT GTT AAC GAT ACC AGC C-3' pYESTrp Reverse 5'-GCG TGA ATG TAA GCG TGA C-3' RVprimer3 5'-CTA GCA AAA TAG GCT GTC CC-3' Rvprimer4 5'-GAC GAT AGT CAT GCC CCG CG-3' GLprimer 1 5'-TGT ATC TTA TGG TAC TGT AAC TG-3' GLprimer 2 5'-CTT TAT GTT TTT GGC GTC TTC CA-3' SeqL-A (ATTL1) 5'-GCG AGA GTA GGG AAC TGC-3'SeqL-B (ATTL2) 5'-AAC ATC AGA GAT TTT GAG ACA C-3' SV40-pArev 5'-CCT CTA CAA ATG TGG TAT GG-3' SV40-Promoter 5'-GCC CCT AAC TCC GCC CAT CC-3'U6 Primer 5'-GGG CAG GAA GAG GGC CTA T-3' Xpress Forward 5'-TAT GGC TAG CAT GAC TGG T-3'1. M13F5’-GGT AAC GCC AGG GTT TTC C-3’2. M13R5’-CAG GAA ACA GCT ATG ACC-3’3. T75’-TAA TAC GAC TCA CTA TAG GG-3’4. T35’-ATT AAC CCT CAC TAA AG-3’5. T7 term.rev5’-TGC TAG TTA TTG CTC AGC GG-3’6. SP65’-GAT TTA GGT GAC ACT ATA G-3’7. KS5’-CTC GAG GTC GAC GGT ATC G-3’8. SK5’-CGC TCT AGA ACT AGT GGA TC-3’测序通用引物序列常见载体的测序引物:Primer of Vector:Vector:Primer(F);Primer(R)pACT T7 T3pACT2 GAL4 AD pACT2-RpAS2-1 GAL4 BD pAS2-1.RpB42AD pB42ADF pB42ADRpBACPAK8 BAC1 BAC2pBK-CMV T7 T3PBS(SK/KS)/M13- M13F/T7 T3/M13RpBV220 PBV220F PBV220RpCAMBIA 1301(1300) P1 P2pCAMBIA 2300 M13R(-48) M13F(-47)PCANTB5E S1/M13R S6pCAT3-enhancer RVP3 此载体无反向引物pcDNA3.0 CMV-F/T7 SP6/BGHpcDNA3.1 T7 BGHpcDNA4 T7/CMV-F BGHpcDNA6 T7/CMV-F(J21025在T7前面)BGHpcDNAII T7 SP6pCE2.1 M13F M13RpCEP4 pCEP-F EBV-RpCF-T M13F M13RpCI T7(17Base)此载体无反向引物pCI-neo T7(17Base)T3pCMS-EGFP T7 T3pCMV-3Tag-4A T3 /PFLAG-CMV-F T7pCMV5 pCMV5F pCMV5RpCMV5-Flag pCMV5F pCMV5RpCMV-MYC/HA pCMV-F pCMV-RpCMV-Sport M13F/T7 SP6/M13RpCMV-Tag(KAN+) T3 T7pCR2.1-TOPO M13F/T7 M13RpCR3.1 T7 BGHpCS2 SP6 T7pDonar M13F M13RpDONR221 M13F M13RpDrive T7 SP6pDRIveR(KAN+) T7 SP6pDsRED1-C1 pDsRED-ex-C1-F pEGFP-N-3’(距离很近,一般不用) pDsRED2-C1(KAN+) pDsRED-ex-C1-F pEGFP-N-3’pDSRED-N1(KAN+) pEGFP-N-5’ PDSRED-N-RpECFP-C pEGFP-C-5’ pECFP-C-3′pECFP-N1 pEGFP-N-5’ pEGFP-N-3’pEF/myc/ER pEF-F pCDNA3.1RpEGFP-C(KAN+) pEGFP-C-5’ pEGFP-C-3′pEGFP-N(KAN+) pEGFP-N-5’ pEGFP-N-3’pENTR/D-TOPO(KAN+) M13F M13RpET-*(His)(KAN+) T7 T7 TerpET22(a,b,c)(AMP+) T7 T7 TerpET28,30,24,49(KAN+) T7 T7 TerpET32(a,b,c)(AMP+) T7/S.tag T7 TerpET50(amp+) T7/s.tag T7TERpET44(AMP+) T7/s.tag T7 TerpEYFP-N1 pEGFP-N-5’ pEGFP-N-3’pEYFP-C1 pEGFP-C-5’ pEGFP-C-3′pFLAG-CMV pFLAG-CMV -F pFLAG-CMV -RpGAD424 GAL4 AD 3′ADpGADGH GAL4 AD 3′ADpGADT7-Rec GAL4 AD/T7 3′ADpGAPZa-A a-FACTOR 3′AOXpGBKT7(Kana+) T7 3′BDpGEM-T(-Easy) M13F/T7 SP6/M13RpGEX-(*)T pGEX-4T-5’ pGEX-4T-3’pGL2 GLP1 GLP2pGL3 RVP3 GLP2(RVP4)(没有特别说明用GLP2)PinpointTM Vector Pinpoint Primer SP6pIRES PIRES-F/T7 T3/PIRES-R (J40720)pIRES2-EGFP pIRES2-EGFP.P5’ pIRES2-EGFP.P3’pLXSN pLXSN-F pLXSN-RpMAL-c2E MalE primer M13F(-47)pMAL-C2x P5’ P3’pMAL-p2X MalE primer M13F(-47)PMD18-T M13R(-48) M13F(-47)PMD19-T M13F(-47) M13R(-48)pPIC9K(AMP+、ZEO+)5’AOX/a-factor 3’AOX pPICZa 5′AOX/PPICZa-F 3′AOXpPROEXHTA M13R(-48) 无反向引物pQE30or40 pQE30-F pQE-R:pRSET T7 T7TERpSilencer3.1-H1 hygro M13F(-47) 3.0REVpSILENCEV2.0-U6 T7 2.0REVpSK01-T M13F M13RpSK-CMV(KAN+) T7 T3pSP72 T7 SP6pSport1 SP6/M13F(-47) T7/M13RpSUPER T7 T3pTA2 M13F/T7(优先使用M13F)T3/M13RpT-Adv M13F /T7 M13RpTARGET TM pTarget.F(在T7前面)/T7 pTarget.R pTO-T7 T7 T7TERpTRC99a-c pTRC99C-F pTRC99C-RpTriPLEx2 5′pTriPLEx2 T7pTWIN-1 T7 T7TER(客户确认再用)pUC18(19)/118(119) M13F(-47) M13R(-48)pUCm-T M13F/T7 M13RpVAX1 T7 BGHpWSK29 T7/M13F/M13F(-47) T3/M13R/M13R(-48)pXT7 T7 SP6pYES2 T7 pYes2.RpLEXA pLEXA-F pLEXA-RpLNCX pLNCX-F pLNCX-RpRECEIVER CMV-F 此载体无反向引物pSHUTTLE-CMV PSHUTTLE-CMV-F PSHUTTLE-CMV-R pSP64/65 SP6 此载体无反向引物pSTBlue-1 T7 SP6pT7Blue(R) T7 M13F(-47)引物名称序列(5′-3′):M13R:CAG GAA ACA GCT ATG ACCM13F:TGT AAA ACG ACG GCC AGTM13F(-47):CGC CAG GGT TTT CCC AGT CAC GACM13R(-48):AGC GGA TAA CAA TTT CAC ACA GGAM13(-96):CCC TCA TAG TTA GCG TAA CGSP6:ATT TAG GTG ACA CTA TAGT7:TAA TAC GAC TCA CTA TAG GGT7 terminator:TGC TAG TTA TTG CTC AGC GGT3:ATT AAC CCT CAC TAA AGG GApGEX-4T-5′:GGG CTG GCA AGC CAC GTT TGG TG pGEX-4T-3′:CCG GGA GCT GCA TGT GTC AGA GGGLp1:TGT ATC TTA TGG TAC TGT AAC TGGLp2:CTT TAT GTT TTT GGC GTC TTC CARVp3:CTA GCA AAA TAG GCT GTC CCRVp4:GAC GAT AGT CAT GCC CCG CGpcDNA3.1R:TAG AAG GCA CAG TCG AGGPinPoint primer:CGT GAC GCG GTG CAG GGC G pCMV-F:TCT AAA AGC TGC GGA ATT GTpCMV-R:TCCAAACTCATCAATGTATCpTRC99C-F: TTG CGC CGA CAT CAT AACpTRC99C-R: CTGCGTTCTGATTTAATCTGpCEP-F: AGA GCT CGT TTA GTG AAC CGEBV-R : GTG GTT TGT CCA AAC TCA TCpIRES2-EGFP.P5’: GTA GGC GTG TAC GGT GGG AG pIRES2-EGFP.P3’: AAC GCA CAC CGG CCT TAT TC3′AD: AGA TGG TGC ACG ATG CAC AGCMV -F:CGC AAA TGG GCG GTA GGC GTGS1:CAA CGT GAA AAA ATT ATT ATT CGCS6:GTA AAT GAA TTT TCT GTA GTA GG5`AOX1:GAC TGG TTC CAA TTG ACA AGC3`AOX1:GCA AAT GGC ATT CTG ACA TCCα-Factor:TAC TAT TGC CAG CAT TGC TGCGAL4 AD:TAC CAC TAC AAT GGA TGpACT2-R:GTGCACGATGCACAGTTGAApB42ADF:CCA GCC TCT TGC TGA GTG GAG ATGpB42ADR:AAG CCG ACA ACC TTG ATT GGA GpEGFP-N-5’:TGG GAG GTC TAT ATA AGC AGA G pEGFP-N-3’:CGT CGC CGT CCA GCT CGA CCA G pEGFP-C-5’:CAT GGT CCT GCT GGA GTT CGT G pEGFP-C-3′ :TAT GGC TGA TTA TGA TCA GTPBV220F:AAG AAG GGC AGC ATT CAA AGPBV220R:CTG CGT TCT GAT TTA ATC TGU6:ATG GAC TAT CAT ATG CTT ACC GTA2.0rev primer:AGG CGA TTA AGT TGG GTA3.0rev:GAG TTA GCT CAC TCA TTA GGCS.tag:GAA CGC CAG CAC ATG GAC5′TriplEx2:CTC CGA GAT CTG GAC GAG CRVP4:GAC GAT AGT CAT GCC CCG CGRVP3:CTA GCA AAA TAG GCT GTC CCpQE30-R:GTT CTG AGG TCA TTA CTG GpQE30-F:TGA GCG GAT AAC AAT TTC ACpEF-F:TCA AGC CTC AGA CAG TGG TTCpDSRED-N1-R:TGA AGC GCA TGA ACT CCT TG pDsRED-express-C1-F:TCC CAC AAC GAG GAC TAC ACpCMV5R:ATT ATA GAG GAC ACC TAG TCpCMV5F:TTC CAA AAT GTC GTA ATA ACP5′:TGC GTA CTG CGG TGA TCA ACP3′:CTG CAA GGC GAT TAA GTT GGBAC2:ACG CAC AGA ATC TAG CGC TTBAC1:AAC CAT CTC GCA AAT AAA TA3′BD:TAA GAG TCA CTT TAA AAT TTG TAT ACpTarget.F:CCA GGA TTT TCC CAG TCA CpTarget.R:GGC TTT ACA CTT TAT GCT TCT7(17base):ACA TCC ACT TTG CCT TTC TCpYes2.R:TCG GTT AGA GCG GAT GTGGAL4 BD:TCA TCG GAA GAG AGT AGpAS2-1.R:AAG AGT TAC TCA AGA ACA AGA ApFlag-CMV-F: GGT AGG CGT GTA CGG TGGpFlag-CMV-R: GCA CTG GAG TGG CAA CTTpIRES-F: CTT ACT GAC ATC CAC TTT GCpIRES.R: CAC TGC ATT CTA GTT GTG GTP1:CCA GGC TTT ACA CTT TAT GCP2:GCG ATT AAG TTG GGT AAC GCpLEXA-F:CGT CAG CAG AGC TTC ACC ATpLEXA-R:TAA AAC CTA AGA GTC ACT TTpLNCX-F:AGC TCG TTT AGT GAA CCG TCA GAT CGpLNCX-R:ACC TAC AGG TGG GGT CTT TCA TTC CCpLXSN-F:CTT GAA CCT CCT CGT TCG ACpLXSN-R:GTT GCT GAC TAA TTG AGA TGpSHUTTLE-CMV-F:GGT CTA TAT AAG CAG AGG TG pSHUTTLE-CMV-R:GTG GTA TGG CTG ATT ATG ATC AG MalE Primer:GGT CGT CAG ACT GTC GAT GAA GCC引物名称序列(5'-3')M13R CAG GAA ACA GCT ATG ACCM13F TGT AAA ACG ACG GCC AGTM13F(-47) CGC CAG GGT TTT CCC AGT CAC GAC M13R(-48) AGC GGA TAA CAA TTT CAC ACA GGA M13(-96) CCC TCA TAG TTA GCG TAA CGSP6 ATT TAG GTG ACA CTA TAGT7 TAA TAC GAC TCA CTA TAG GGT7 terminator TGC TAG TTA TTG CTC AGC GGT3 ATT AAC CCT CAC TAA AGG GA pGEX-4T-5' GGG CTG GCA AGC CAC GTT TGG TG pGEX-4T-3' CCG GGA GCT GCA TGT GTC AGA GG GLp1 TGT ATC TTA TGG TAC TGT AAC TG GLp2 CTT TAT GTT TTT GGC GTC TTC CA RVp3 CTA GCA AAA TAG GCT GTC CCRVp4 GAC GAT AGT CAT GCC CCG CG pcDNA3.1R TAG AAG GCA CAG TCG AGG PinPoint primer CGT GAC GCG GTG CAG GGC G pCMV-F TCT AAA AGC TGC GGA ATT GT pCMV-R TCCAAACTCATCAATGTATCpTRC99C-F: TTG CGC CGA CAT CAT AACpTRC99C-R: CTGCGTTCTGATTTAATCTGpCEP-F: AGA GCT CGT TTA GTG AAC CG EBV-R : GTG GTT TGT CCA AAC TCA TC pIRES2-EGFP.P5’: GTA GGC GTG TAC GGT GGG AG pIRES2-EGFP.P3’: AAC GCA CAC CGG CCT TAT TC3'AD: AGA TGG TGC ACG ATG CAC AG CMV -F CGC AAA TGG GCG GTA GGC GTGS1 CAA CGT GAA AAA ATT ATT ATT CGC S6 GTA AAT GAA TTT TCT GTA GTA GG 5`AOX1 GAC TGG TTC CAA TTG ACA AGC3`AOX1 GCA AAT GGC ATT CTG ACA TCCα-Factor TAC TAT TGC CAG CAT TGC TGC GAL4 AD TAC CAC TAC AAT GGA TGpACT2-R GTGCACGATGCACAGTTGAApB42ADF: CCA GCC TCT TGC TGA GTG GAG ATG pB42ADR: AAG CCG ACA ACC TTG ATT GGA G pEGFP-N-5’: TGG GAG GTC TAT ATA AGC AGA G pEGFP-N-3’: CGT CGC CGT CCA GCT CGA CCA G pEGFP-C-5’: CAT GGT CCT GCT GGA GTT CGT G pEGFP-C-3' : TAT GGC TGA TTA TGA TCA GTPBV220F: AAG AAG GGC AGC ATT CAA AG PBV220R: CTG CGT TCT GAT TTA ATC TGU6 ATG GAC TAT CAT ATG CTT ACC GTA2.0rev primer: AGG CGA TTA AGT TGG GTA3.0rev: GAG TTA GCT CAC TCA TTA GGC S.tag GAA CGC CAG CAC ATG GAC5'TriplEx2 CTC CGA GAT CTG GAC GAG CRVP4 GAC GAT AGT CAT GCC CCG CG RVP3 CTA GCA AAA TAG GCT GTC CC pQE30-R GTT CTG AGG TCA TTA CTG GpQE30-F TGA GCG GAT AAC AAT TTC ACpEF-F TCA AGC CTC AGA CAG TGG TTC pDSRED-N1-R TGA AGC GCA TGA ACT CCT TG pDsRED-express-C1-F TCC CAC AAC GAG GAC TAC ACpCMV5R ATT ATA GAG GAC ACC TAG TCpCMV5F TTC CAA AAT GTC GTA ATA ACP5' TGC GTA CTG CGG TGA TCA ACP3' CTG CAA GGC GAT TAA GTT GGBAC2 ACG CAC AGA ATC TAG CGC TTBAC1 AAC CAT CTC GCA AAT AAA TA3'BD: TAA GAG TCA CTT TAA AAT TTG TAT AC pTarget.F CCA GGA TTT TCC CAG TCA C pTarget.R GGC TTT ACA CTT TAT GCT TCT7(17base) ACA TCC ACT TTG CCT TTC TCpYes2.R TCG GTT AGA GCG GAT GTGGAL4 BD TCA TCG GAA GAG AGT AGpAS2-1.R AAG AGT TAC TCA AGA ACA AGA A pFlag-CMV-F: GGT AGG CGT GTA CGG TGGpFlag-CMV-R: GCA CTG GAG TGG CAA CTTpIRES-F: CTT ACT GAC ATC CAC TTT GC pIRES.R: CAC TGC ATT CTA GTT GTG GTP1 CCA GGC TTT ACA CTT TAT GCP2 GCG ATT AAG TTG GGT AAC GC pLEXA-F CGT CAG CAG AGC TTC ACC AT pLEXA-R TAA AAC CTA AGA GTC ACT TT pLNCX-F AGC TCG TTT AGT GAA CCG TCA GAT CG pLNCX-R ACC TAC AGG TGG GGT CTT TCA TTC CC pLXSN-F CTT GAA CCT CCT CGT TCG AC pLXSN-R GTT GCT GAC TAA TTG AGA TG pSHUTTLE-CMV-F GGT CTA TAT AAG CAG AGG TG pSHUTTLE-CMV-R GTG GTA TGG CTG ATT ATG ATC AG MalE Primer GGT CGT CAG ACT GTC GAT GAA GCC引物名称 引物序列3’AOX 5'-GGC AAA TGG CAT TC TGA CAT-3'3'AD 5'-AGA TGG TGC ACG ATG CAC AG-3'3'BD 5'-TAA GAG TCA CTT TAA AAT TTG TAT C-3'5’AOX1 5'-GAC TGG TTC CAA TTG ACA AGC-3'A-FACTOR 5'-TAC TAT TGC CAG CAT TGC TGC-3'BAC1 5'-AAC CAT CTC GCA AAT AAA TA-3'BAC2 5'-ACG CAC AGA ATC TAG CGC TT-3'BGH rev 5'-TAG AAG GCA CAG TCG AGG C-3'CMV-F 5'-CGC AAA TGG GCG GTA GGC GTG-3'CMV-R 5'-GTT CAC GGT GCC CTC C-3'DsRed1-C-r 5'-AGC TGG ACA TCA CCT CCC ACA ACG-3'DsRed1-N-f 5'-GTA CTG GAA CTG GGG GGA CAG-3'EBV Reverse 5'-TGT GGT TTG TCC AAA CTC AAT C-3' EBV-R 5'-GTG GTT TGT CCA AAC TCA TC-3'GAL4 AD-F 5'-TAC CAC TAC AAT GGA TG-3'GAL4 BD 5'-TCA TCG GAA GAG AGT AG-3'GLP1 5'-TGT ATC TTA TGG TAC TGT AAC TG-3' GLP2 5'-CTT TAT GTT TTT GGC GTC TTC CA-3'M13(-96) 5'-CCC TCA TAG TTA GCG TAA CG-3'M13F 5'-TGT AAA ACG ACG GCC AGT-3'M13F(-47)5'-CGC CAG GGT TTTC CCA GTC ACG AC-3' M13R 5'-CAG GAA ACA GCT ATG ACC-3'M13R(-48)5'-AGC GGA TAA CAA TTT CAC ACA GGA-3' P24990 5'-TCATCGGAAGCGAGTAG-3'P31430 5'-CGT TTT AAAACCTAAGAGTCAC-3'P12584 5'-TTT TCA GTA TCT ACG AT-3’P17110 5'-TAC CAC TAC AAT GGA TG-3’PAS2-1F 5'-TCA TCG GAA GCG AGT AG-3'PAS2-1R 5'-CGT TTT AAA ACC TAA GAG TCA C-3'PB42ADF 5'-CCA GCC TCT TGC TGA GTG GAG ATG-3' PB42ADR 5'-AAG CCG ACA ACC TTG ATT GGA G-3'pbd-gal4-f 5'-GCC TCT AAC ATT GAG ACA GC-3'pbd-gal4-r 5'-AAG AGT TAC TCA AGA ACA AGA A-3'pbi-121(35s) 5'-GAC GCA CAA TCC CAC TAT CC-3'PBV220F 5'-AAG AAG GGC AGC ATT CAA AG-3'PBV220R 5'-CTG CGT TCT GAT TTA ATC TG-3' PCDNA3.0-F2 5'-CTC TGG CTA ACT AGA GAA CC-3' PCDNA3.0-R2 5'-GGC AAC TAG AAG GCA CAG TC-3' PCDNA3.1F 5'-CTA GAG AAC CCA CTG CTT AC-3’ PCDNA3.1R 5'-TAG AAG GCA CAG TCG AGG-3’pCEP Frorward 5'-AGC AGA GCT CGT TTA GTG AAC CG-3' PCEP-F 5'-AGA GCT CGT TTA GTG AAC CG-3'PCMV-F 5'-TCT AAA AGC TGC GGA ATT GT-3'PCMV-F 5'-TTC CAA AAT GTC GTA ATA AC-3'PCMV-R 5'-TCC AAA CTC ATC AAT GTA TC-3'PCMV-R 5'-ATT ATA GAG GAC ACC TAG TC-3' PDONR-F 5'-TCG CGT TAA CGC TAG CAT GGA TCT C-3' PDONR-R 5'-GTA ACA TCA GAG ATT TTG AGA CAC-3’ PEF-F 5'-TCA AGC CTC AGA CAG TGG TTC-3' PEGFP-C-3’ 5'-TAT GGC TGA TTA TGA TCA GT-3' PEGFP-C-5’ 5'-CAT GGT CCT GCT GGA GTT CGT G-3' PEGFP-N-3’ 5'-CGT CGC CGT CCA GCT CGA CCA G-3' PEGFP-N-5’ 5'-TGG GAG GTC TAT ATA AGC AGA G-3' pETup 5'-ACG ATG CGT CCG GCG TAG AGG A-3' PGEX-3’ 5'-CCG GGA GCT GCA TGT GTC AGA GG-3' PGEX-5’ 5'-GGG CTG GCA AGC CAC GTT TGG TG-3' PIRES2-EGFP-F 5'-GTA GGC GTG TAC GGT GGG AG-3'PIRES3-EGFP-R 5'-AAC GCA CAC CGG CCT TAT TC-3'pMAL-C2X-F 5'-TGC GTA CTG CGG TGA TCA AC-3’pMAL-C2X-R 5'-CTG CAA GGC GAT TAA GTT GG-3’PQE30F 5'-TGA GCG GAT AAC AAT TTC AC-3'PQE30R 5'-GTT CTG AGG TCA TTA CTG G-3' pTARGET-SEQ 5'-TTA CGC CAA GTT ATT TAG GTG ACA-3’ PTRC99C-F 5'-TTG CGC CGA CAT CAT AAC-3’PTRC99C-R 5'-CTG CGT TCT GAT TTA ATC TG-3’ PTRCHISA-F 5'-CAT GGT ATG GCT AGC ATG AC-3’ PTRCHISA-R 5'-CAG GCT GAA AAT CTT CTC TC-3’RVP3 5'-CTA GCA AAA TAG GCT GTC CC-3'RVP4 5'-GAC GAT AGT CAT GCC CCG CG-3'S.TAG 5'-CGA ACG CCA GCA CAT GGA CA-3'S1 5'-CAA CGT GAA AAA ATT ATT ATT CGC-3’S6 5'-GTA AAT GAA TTT TCT GTA TGA GG-3’ SP6 5'-ATT TAG GTG ACA CTA TAG-3'T3 5'-ATT AAC CCT CAC TAA AGG GA-3'T7 5'-TAA TAC GAC TCA CTA TAG GG-3'T7(17BASE)5'-ACA TCC ACT TTG CCT TTC TC-3'T7TER 5'-TGC TAG TTA TTG CTC AGC GG-3'TRXF 5'-CAT ATG AGC GAT AAA ATT ATT CAC-3' TRXR 5'-GGA TCC CTT GTC ATC GTC ATC ACC A-3' malEfor 5'-GGT CGT CAG ACT GTC GAT GAA GCC-3' pTrcHis-fw 5'-TTA AAG AGG TAT ATA TTA ATG TAT CG-3' Xpressfor 5'-AGC ATG ACT GGT GGA CAG-3'pTrcHis-rv 5'-ATC TGT ATC AGG CTG AAA ATC-3'N-CMV-30 5'-ATA ACC CCG CCC CGT TG-3'C-CMV-24 5'-TTA GGA CAA GGC TGG TGG-3'CMV-Profor 5'-ATG GGC GGT AGG CGT G-3'pCAMBIA1301-5'(ECORI) 5'-CCA GGC TTT ACA CTT TAT GC-3' pCAMBIA1301-3'(HindIII) 5'-GCG ATT AAG TTG GGT AAC GC-3'T7 EEV 5'-ATG TCG TAA TAA CCC CGC CCC G-3' pDBLeu-5' 5'-GAA TAA GTG CGA CAT CAT CAT C- 3’ pDBLeu-3' 5'-CAG ATG GAA CGG TCT TTA AAT G-3’ HGHrev 5'-CTG GGG AGG GGT CAC AGG G-3'PSG5(BAMHI)-F 5'-GCT GGT TAT TGT GCT GTC TC-3'PSG5-R(SV40) 5'-AAA CCA CAA CTA GAA TGC AGT-3'M13R(-26) 5'-CAG GAA ACA GCT ATG AC-3'pBRforBam 5'-CTT GGA GCC ACT ATC GAC-3' pBRrevBam 5'-GGT GAT GTC GGC GAT ATA GG-3' pBRforEco 5'-AAT AGG CGT ATC ACG AGG C-3' pBRrevHind 5'-GCG TTA GCA ATT TAA CTG TG-3'c-mycrev 5'-CTC TTC TGA GAT GAG TTT TTG-3'pGAL4-ADrv 5'-GAA CTT GCG GGG TTT TTC-3'pGAL4-BDrv 5'-AAT CAT AAG AAA TTC GCC C-3'pMD19T490R 5'-CACAGGAAACAGCTATGACC-3'Psos5' 5'-CCA AGA CCA GGT ACC ATG-3'Psos3' 5'-GCC AGG GTT TTC CCA GT-3'Primer TriplEx-LD-5´ 5'-CTC GGG AAG CGC GCC ATT GTG TTG G-3'Primer TriplEx-LDrv-3' 5'-ATA CGA CTC ACT ATA GGG CGA ATT GGC-3' UB-F 5'-TCA GTG TTA GAC TAG TAA ATT-3'27F 5'-AGA GTT TGA TCC TGG CTC AG-3’1492R 5'-GGY TAC CTT GTT ACG ACT T-3’PTR5 5'-CTC CGA GAT CTG GAC GAG C-3'pLXSN-F CTT GAA CCT CCT CGT TCG ACpLXSN-R GTT GCT GAC TAA TTG AGA TGpLEXA-F CGT CAG CAG AGC TTC ACC ATpLEXA-R TAA AAC CTA AGA GTC ACT TTpLNCX-F AGC TCG TTT AGT GAA CCG TCA GAT CG pLNCX-R ACC TAC AGG TGG GGT CTT TCA TTC CC pDSRED-N-R TGA AGC GCA TGA ACT CCT TGmtDNA-D-LoopF AGG ACT Acg gct tga aaa gcmtDNA-D-LoopR atg tgc ctg acc gag gaa cca gH1primer CGCTATGTGTTCTGGGAAATpRECEIVER-M03F 5'-GCGGTAGGCGTGTACGGT-3'pRECEIVER-M03R 5'-CCGGACACGCTGAACTTGT-3'ACYCDuetUP1Primer #71178-3 GGATCTCGACGCTCTCCCTDuetUP2 Primer TTGTACACGGCCGCATAATCDuetDOWN1 GATTATGCGGCCGTGTACAA1525R 5'-AgAAAggAggTgATCcAgcc-3'pBAD_F ATGCCATAGCATTTTTATCCpBAD_R GATTTAATCTGTATCAGGAUG1F 5'-CAATTTACATCTTTATTTATTAACG-3'AUG1R 5'-GAAGAGAAAAACATTAGTTGGC-3'V5rev 5'-ATC GAG ACC GAG GAG AGG -3'EGFP-Nrev 5'-CGT CGC CGT CCA GCT C-3'TKPAreverse 5΄-CTTCCGTGTTTCAGTTAGC-3΄pDsRED-express-C1-F TCC CAC AAC GAG GAC TAC ACDEST22F 5’-tataacgcgtttggaatcact-3’DEST22R 5’-agccgacaaccttgattggagac-3’pBMN_5' gcttggatacacgccgcpYes2.R TCG GTT AGA GCG GAT GTGpSG5-3 GAAATTTGTGATGCTATTGCpMSCV-5' CCCTTGAACCTCCTCGTTCGACCpMSCV-3' GAGACGTGCTACTTCCATTTGTC内参通用引物 16SDNAsprimer sequence27F 5’-AGAGTTTGATCCTGGCTCAG-3’515F 5’-GTGCCAGCMGCCGCGGTAA-3’21F 5’-TTCCGGTTGATCCTGCCGGA-3’312F 5’-TCCTACGGGAGGCAGCAGCT-31492R 5’-GGYTACCTTGTTACGACTT-3’ 1525R 5'-AgAAAggAggTgATCcAgcc-3' ITS1 TCCGTAGGTGAACCTGCGGITS4 TCCTCCGCTTATTGATATGC真菌通用引物?nu-SSU-0817-5’ TTAGCATGGAATAATRRAATAGGAnu-SSU-1536-3’ ATTGCAATGCYCTATCCCCA。