SuperScript™ III Reverse Transcriptase
Description:
SuperScript™
III Reverse Transcriptase (RT) is a
proprietary mutant of SuperScript™
II RT that is active at 50°C
and has a half-life of 220 minutes,
providing increased specificity with
Gene-Specific Primers (GSPs) and the highest
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SuperScript™
III Reverse Transcriptase (RT) is a
proprietary mutant of SuperScript™
II RT that is active at 50°C and has a
half-life of 220 minutes, providing
increased specificity with Gene-Specific
Primers (GSPs) and the highest cDNA yield of
all RTs. It is ideal for RT-PCR of a
specific gene or generating cDNA from total
or poly (A)+ RNA sample. Like SuperScript™
II, it synthesizes a complementary DNA
strand from single-stranded RNA, DNA, or an
RNA:DNA hybrid. SuperScript™
III RT is genetically engineered by the
introduction of point mutations that
increase half-life, reduce RNase activity,
and increase thermal stability. The
structural modifications provide:
-
Half life of 220 minutes at 50°C
for the highest cDNA yields (Figure 1)
-
Reduced RNase H activity for more
full-length cDNA
-
Full activity at 50°C
for increased specificity with GSP
-
Ability to increase RT units without
inhibiting subsequent PCR
Applications:
Synthesis of first-strand cDNA, array
labeling, cDNA libraries, RT-PCR, primer
extension, and 3´
and 5´ RACE.
Source:
Purified from E. coli expressing the
pol gene of M-MLV (1,2), mutagenized
to increase thermal stability and half life
and reduce RNase H activity.
Performance and Quality Testing:
SDS-PAGE purity; endodeoxyribonuclease,
exodeoxyribonuclease, and ribonuclease
assays (Figure 2) and yield and length of
cDNA product.
Unit Definition:
One unit of SuperScript™
III RT is the amount of enzyme required to
incorporate 1 nmole of deoxyribonucleotide
into acid-precipitable material in 10 min.
at 37°C using poly(A)•oligo(dT)12-18
as a template•primer
(3).
Unit Reaction Conditions:
50 mM Tris-HCl (pH 8.3), 40 mM KCl, 6 mM
MgCl 2 , 1 mM DTT. 0.5 mM [ 3 H]dTTP, 0.1 mM
poly(A), 0.1 mM oligo(dT)12-18 ,
0.1 mg/ml BSA, and enzyme in 50 µl for 10
min. at 37°C.
Contents and Storage:
SuperScript™
III RT is supplied with a vial of 5X
first-strand buffer [250 mM Tris-HCl (pH
8.3), 375 mM KCl, 15 mM MgCl2],
and a vial of 100 mM DTT. Store at -20°C.
Guaranteed stable for 6 months when properly
stored
Ordering Information
|
名称 |
规格 |
报价 |
折后价 |
货号 |
产地 |
|
SuperScript™ III Reverse Transcriptase |
10000 units |
3604 |
3263现货 |
18080-044 |
Invitrogen |
