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TOPscript™ cDNA synthesis Kit (mix type/dN6 plus)
Product
description
TOPscript™ cDNA synthesis Kit (mix type/dN6 plus)
is designed for the maximum-efficiency conversion of all types of RNA including
mRNA to cDNA. The kit contains RNase inhibitor in TOPscript™ Reverse TranscriptaseR+ to protect
RNA degradation and random hexamer (dN6) in 10X TOPscript™ RT reaction MIX (dN6). TOPscript™ Reverse Transcriptase is a highly thermostable
version of M-MLV RT (RNase H-), and it can synthesize cDNA at 50~60℃ with highest fidelity. Thus, it is well suited
to synthesize long cDNA (>20 kb) from long and highly secondary-structured
RNA.
Characteristics
- cDNA
synthesis across a broad temperature range
- Synthesis
long cDNA
- Include
all components required for cDNA synthesis
Applications
-
First-strand of cDNA synthesis
- Primer
extension
Supplied
with
- TOPscript™ Reverse TranscriptaseR+ (200 U/μl)
- 10X TOPscript™ RT reaction MIX (dN6)
- Sterile water (RNase free)
Quality
control
- Purity:
>99% on SDS-PAGE
-
Endonuclease-free
-
Exonuclease-free
- RNase-free
-
Inhibitor-free
-
Satisfactory yield and length of cDNA products
Unit
definition
One unit is
defined as the amount of enzyme required to incorporate 1 nmol of dTTP into
acid-insoluble materrials using 0.4 mM poly(rA)-oligo(dT) as substrate at 37℃ in 10 min in a 1X reaction buffer.
Storage
buffer
50 mM
tris-HCl (pH 7.6), 150 mM NaCl, 1mM dTT, 0.1 mM EDTA, 0.1% NP-40, 50% glycerol
Standard
reaction conditions
10X TOPscript™ RT reaction
MIX (dN6) |
2 μl
|
TOPscript™
Reverse TransciptaseR+ (200 U/μl) |
1 μl |
a)Template RNA |
X μl |
Sterile water (RNase free) |
up
to 20 μl |
a)Prepare
one of the following RNA template.
- Total RNA: 1 ng~5 μg
-
Messenger RNA (mRNA): 1 ng~250 ng
- Specific RNA: 0.01 pg~0.5 μg
→ Incubate at 42℃~60℃ for 60 min.
→ Incubate at 95℃ for 5 min to inactivate the reaction.
Note
TOPscript™ Reverse Transcriptase performs optionally over
the full range of 42℃-60℃. Typically, 50℃
is a good starting point. For RNAs containing secondary structure and other
challenging targets, a synthesis temperature of 60℃ may be used without loss of performance.
● Material Safety Data Sheet
- Enzynomics_MSDS_EZ205_TOPscript™ cDNA synthesis Kit (mix type/dN6 plus)_E