逆转录试剂盒( transcriptor first strand cdna synthesis kit ) 说明书
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Ϫ70°C. N Avoid repeated freezing and thawing.
• standard laboratory equipment – nuclease-free, aerosol-resistant pipette tips – pipettes with disposable, positive-displacement tips – sterile reaction tubes for preparing master mixes and dilutions – standard benchtop microcentrifuge – thermal block cycler with a heated lid – sequence-specific PCR primers (optional)
L In Cat. No. 04 896 866 001 and Cat. No. 04 897 030 001 the control reagents (vial 7 and 8) are not included. Therefore, in these kits vial 7 is Water, PCR Grade.
C O
L
PCR in a conventional thermal block cycler
15 L
PCR on the LightCycler® Carousel-Based System
16
PCR on the LightCycler® 480 Instrument
18
3. Results.................................................................................................................................... 20
Transcriptor Reverse Transcriptase
a) 1 vial, 25 l (20 U/l) b) 1 vial, 50 l (20 U/l) c) 2 vials, each 50 l (20 U/l) • Storage buffer: 200 mM potassium phosphate,
Transcriptor First Strand cDNA Synthesis Kit
y Version 6.0
1. What this Product Does
Number of Tests The kit is designed for 50, 100 or 200 reactions (depending on pack size).
www.roche-applied-science.com
3
04 379 012 001
Transcriptor First Strand cDNA Synthesis Kit y Version 6.0
1. What this Product Does, continued
Vial/ Cap
Label
4. Troubleshooting.................................................................................................................... 23
5. Additional Information on this Product........................................................................... 26
c) 2 vials, each 100 l (40 U/l)
• Storage buffer: 20 mM Hepes-KOH, 50 mM
KCl, 8 mM dithiothreitol, 50% glycerol (v/v),
pH approx. 7.6 (at 4°C)
Deoxynuc-leo- a) 1 vial, 100 l (yellow cap)
O
Procedure B: cDNA Synthesis with anchored-oligo(dT)18 primer AND random hexamer primer 11 O
来自百度文库
C O
2.3 Control Reaction cDNA Synthesis PCR for PBGD
14 14 15
tide Mix
b) 1 vial, 200 l (purple cap)
c) 2 vials, each 200 l (purple cap)
• 10 mM each dATP, dCTP, dGTP, dTTP
Anchoredoligo(dT)18 Primer
a) 1 vial, 100 l (50 M) b) 1 vial, 200 l (50 M) c) 2 vials, each 200 l (50 M)
• 5ϫ conc.: 250 mM Tris/HCl, 150 mM KCl,
40 mM MgCl2, pH approx. 8.5 (25°C)
Protector
a) 1 vial, 50 l (40 U/l)
RNase Inhibitor b) 1 vial, 100 l (40 U/l)
For life science research only. Not for use in diagnostic procedures.
Transcriptor First Strand cDNA Synthesis Kit
Cat. No. 04 379 012 001
Cat. No. 04 896 866 001 Cat. No. 04 897 030 001
Storage and Stability
Additional Equipment and Reagents Required
Application
Store the kit at -15 to -25°C through the expiration date printed on the label. L The Control RNA (vial 7 in Cat. No. 04 379 012 001) should be stored at
fraction purified from an immortalized cell line (K562)
Control Primer a) 1 vial, 40 l
Mix PBGD
• 5 M forward and reverse primer specific for
human porphobilinogen deaminase (PBGD)
Kit for 50 reactions including 10 control reactions Kit for 100 reactions Kit for 200 reactions
www.roche-applied-science.com
Table of Contents
1. What this Product Does ........................................................................................................ 3
9 (7 for b,c) colorless
Water, PCR-grade
Content
a) Cat. No. 04 379 012 001 b) Cat. No. 04 896 866 001 c) Cat. No. 04 897 030 001
a) 1 vial, 1 ml b) 2 vials, each 1ml c) 3 vials, each 1 ml
Kit Contents
Vial/ Cap
1 red
2 colorless
3 colorless
4 yellow/ purple
5 blue
6 blue
7 green
8 green
Label
Content
a) Cat. No. 04 379 012 001 b) Cat. No. 04 896 866 001 c) Cat. No. 04 897 030 001
Random Hexamer Primer
a) 1 vial, 100 l (600 M) b) 1 vial, 200 l (600 M) c) 2 vials, each 200 l (600 M)
Control RNA
a) 1 vial, 20 l (50 ng/l) • contains a stabilized solution of a total RNA
2.1 Before You Begin
5
Precautions
5
P
Sample Material Primer
R 2.2 Procedure
5 6
P
8R
O
Standard RT-PCR Procedure
8O
T
Procedure A: cDNA Synthesis with anchored-oligo(dT)18 OR random OR sequence-specific primer 8 T
6.1 Conventions
29
Text Conventions
29
Symbols
29
6.2 Changes to Previous Version
29
6.3 Ordering Information
29
6.4 Trademarks
31
2 04 379 012 001
www.roche-applied-science.com
* available from Roche Applied Science
Store the kit at ؊15 to ؊25°C N Store Control RNA (vial 7 in
Cat. No 04 379 012 001) at —70°C or below.
y Version 6.0
Content version: September 2010
2 mM dithiothreitol, 0.2% Triton X-100 (v/v),
50% glycerol (v/v), pH approx. 7.2
Transcriptor RT a) 1 vial, 1 ml
Reaction Buf- b) 1 vial, 1 ml
fer (5ϫ)
c) 2 vials, each 1 ml
• for control reactions in combination with a LightCycler® Instrument: – LightCycler® 1.5 Instrument*, LightCycler® 2.0 Instrument* or LightCycler® 480 Instrument*
How this Product Works
26
References
27
Quality Control
28
6. Supplementary Information............................................................................................... 29
Number of Tests
3
Kit Contents
3
Storage and Stability
4
Additional Equipment and Reagents Required
4
Application
4
2. How to Use this Product....................................................................................................... 5
• standard laboratory equipment – nuclease-free, aerosol-resistant pipette tips – pipettes with disposable, positive-displacement tips – sterile reaction tubes for preparing master mixes and dilutions – standard benchtop microcentrifuge – thermal block cycler with a heated lid – sequence-specific PCR primers (optional)
L In Cat. No. 04 896 866 001 and Cat. No. 04 897 030 001 the control reagents (vial 7 and 8) are not included. Therefore, in these kits vial 7 is Water, PCR Grade.
C O
L
PCR in a conventional thermal block cycler
15 L
PCR on the LightCycler® Carousel-Based System
16
PCR on the LightCycler® 480 Instrument
18
3. Results.................................................................................................................................... 20
Transcriptor Reverse Transcriptase
a) 1 vial, 25 l (20 U/l) b) 1 vial, 50 l (20 U/l) c) 2 vials, each 50 l (20 U/l) • Storage buffer: 200 mM potassium phosphate,
Transcriptor First Strand cDNA Synthesis Kit
y Version 6.0
1. What this Product Does
Number of Tests The kit is designed for 50, 100 or 200 reactions (depending on pack size).
www.roche-applied-science.com
3
04 379 012 001
Transcriptor First Strand cDNA Synthesis Kit y Version 6.0
1. What this Product Does, continued
Vial/ Cap
Label
4. Troubleshooting.................................................................................................................... 23
5. Additional Information on this Product........................................................................... 26
c) 2 vials, each 100 l (40 U/l)
• Storage buffer: 20 mM Hepes-KOH, 50 mM
KCl, 8 mM dithiothreitol, 50% glycerol (v/v),
pH approx. 7.6 (at 4°C)
Deoxynuc-leo- a) 1 vial, 100 l (yellow cap)
O
Procedure B: cDNA Synthesis with anchored-oligo(dT)18 primer AND random hexamer primer 11 O
来自百度文库
C O
2.3 Control Reaction cDNA Synthesis PCR for PBGD
14 14 15
tide Mix
b) 1 vial, 200 l (purple cap)
c) 2 vials, each 200 l (purple cap)
• 10 mM each dATP, dCTP, dGTP, dTTP
Anchoredoligo(dT)18 Primer
a) 1 vial, 100 l (50 M) b) 1 vial, 200 l (50 M) c) 2 vials, each 200 l (50 M)
• 5ϫ conc.: 250 mM Tris/HCl, 150 mM KCl,
40 mM MgCl2, pH approx. 8.5 (25°C)
Protector
a) 1 vial, 50 l (40 U/l)
RNase Inhibitor b) 1 vial, 100 l (40 U/l)
For life science research only. Not for use in diagnostic procedures.
Transcriptor First Strand cDNA Synthesis Kit
Cat. No. 04 379 012 001
Cat. No. 04 896 866 001 Cat. No. 04 897 030 001
Storage and Stability
Additional Equipment and Reagents Required
Application
Store the kit at -15 to -25°C through the expiration date printed on the label. L The Control RNA (vial 7 in Cat. No. 04 379 012 001) should be stored at
fraction purified from an immortalized cell line (K562)
Control Primer a) 1 vial, 40 l
Mix PBGD
• 5 M forward and reverse primer specific for
human porphobilinogen deaminase (PBGD)
Kit for 50 reactions including 10 control reactions Kit for 100 reactions Kit for 200 reactions
www.roche-applied-science.com
Table of Contents
1. What this Product Does ........................................................................................................ 3
9 (7 for b,c) colorless
Water, PCR-grade
Content
a) Cat. No. 04 379 012 001 b) Cat. No. 04 896 866 001 c) Cat. No. 04 897 030 001
a) 1 vial, 1 ml b) 2 vials, each 1ml c) 3 vials, each 1 ml
Kit Contents
Vial/ Cap
1 red
2 colorless
3 colorless
4 yellow/ purple
5 blue
6 blue
7 green
8 green
Label
Content
a) Cat. No. 04 379 012 001 b) Cat. No. 04 896 866 001 c) Cat. No. 04 897 030 001
Random Hexamer Primer
a) 1 vial, 100 l (600 M) b) 1 vial, 200 l (600 M) c) 2 vials, each 200 l (600 M)
Control RNA
a) 1 vial, 20 l (50 ng/l) • contains a stabilized solution of a total RNA
2.1 Before You Begin
5
Precautions
5
P
Sample Material Primer
R 2.2 Procedure
5 6
P
8R
O
Standard RT-PCR Procedure
8O
T
Procedure A: cDNA Synthesis with anchored-oligo(dT)18 OR random OR sequence-specific primer 8 T
6.1 Conventions
29
Text Conventions
29
Symbols
29
6.2 Changes to Previous Version
29
6.3 Ordering Information
29
6.4 Trademarks
31
2 04 379 012 001
www.roche-applied-science.com
* available from Roche Applied Science
Store the kit at ؊15 to ؊25°C N Store Control RNA (vial 7 in
Cat. No 04 379 012 001) at —70°C or below.
y Version 6.0
Content version: September 2010
2 mM dithiothreitol, 0.2% Triton X-100 (v/v),
50% glycerol (v/v), pH approx. 7.2
Transcriptor RT a) 1 vial, 1 ml
Reaction Buf- b) 1 vial, 1 ml
fer (5ϫ)
c) 2 vials, each 1 ml
• for control reactions in combination with a LightCycler® Instrument: – LightCycler® 1.5 Instrument*, LightCycler® 2.0 Instrument* or LightCycler® 480 Instrument*
How this Product Works
26
References
27
Quality Control
28
6. Supplementary Information............................................................................................... 29
Number of Tests
3
Kit Contents
3
Storage and Stability
4
Additional Equipment and Reagents Required
4
Application
4
2. How to Use this Product....................................................................................................... 5