FicoScript®
Reverse Transcriptase is an engineered version of M-MLV
RT with reduced RNase H activity and increased thermal
stability. The enzyme is purified to near homogeneity
from E.
coli containing
the modified pol
gene
of Moloney Murine Leukemia Virus . The enzyme can be
used to synthesize first-strand cDNA at temperatures up
to 48°C,
providing increased specificity, higher yields of cDNA,
and more fulllength product than other reverse
transcriptases. It can generate cDNA from 100 bp to
>12 kb.It is suitable for qPCR.
1
、
Enzyme Storage
Buffer: M-MLV
Reverse Transcriptase is supplied in 20mM Tris-HCl (pH
7.5), 200mM NaCl, 0.1mM EDTA, 1mM DTT, 0.01% Nonidet
P-40 and 50% glycerol.
2
、
M-MLV Reverse
Transcriptase 5X Reaction Buffer: When
the M-MLV Reverse Transcriptase 5X Reaction Buffer
supplied with this enzyme is diluted 1:5, it has a
composition of 50mM Tris-HCl (pH 8.3 25°C ), 75mM KCl,
3mM MgCl 2 and 10mM DTT.
3
、
Source: Purified
from an E. coli strain expressing a recombinant clone.
4
、
Storage Conditions: Store
at – 20°C . Avoid exposure to frequent temperature
changes.
5
、
Unit Definition: One
unit is defined as the amount of enzyme required to
catalyze the transfer of 1nmol of deoxynucleotide into
acid-precipitable material in 10 minutes at 37°C . The
reaction conditions are: 50mM Tris-HCl (pH 8.3), 7mM
MgCl 2 ,
6
、
Usage Note: M-MLV
Reverse Transcriptase is less processive than AMV
Reverse Transcriptase, and therefore, more units of the
M-MLV enzyme are required to generate the same amount of
cDNA as in the AMV reaction. Thus, starting with 1μg of
mRNA in a first-strand cDNA synthesis, 200 units of the
M-MLV enzyme are recommended as opposed to 25 units of
the AMV enzyme.
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Reserved.
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