タイトルまたは説明
M-MLV GIII Reverse Transcriptase (third generation
enzyme)
Cat. No.: M***4
Price: $*0 for ****0U (negotiable if bulk purchase)
Storage conditions: **0℃
Product composition
|
Component
specification
|
Specification
|
|
M-MLVGIII Reverse Transcriptase(**0U/ul)
|
*0ul
|
|
5×M-MLV First Strand Buffer
|
**0ul
|
|
0.1M DTT
|
**0ul
|
Product Introduction
M-MLV GIII Reverse Transcriptase is a third-generation M-MLV
reverse transcriptase obtained through gene modification and
recombinant technology. Compared with wild-type M-MLV reverse
transcriptase, this enzyme removes RNase H activity and greatly
improves thermal stability (optimal reaction temperature is *0°C),
thereby greatly improving the specificity and length of the
first-chain cDNA synthesis (up to *2 kb), and enhancing tolerance
to complex secondary structures of RNA.
Enzyme activity unit definition
At *7°C, with Poly(A)-Oligo(dT) as template/primer, within *0 min,
the amount of enzyme required to incorporate 1 nmol of [3H] dTTP is
defined as 1 activity unit (U).
Quality Control
Host DNA detection
No host DNA contamination
Endonuclease residue detection
The enzyme solution was incubated with supercoiled plasmid DNA at
*7°C for 4 hours, and the plasmid was detected by DNA
electrophoresis without any changes.
Exonuclease residue detection
Incubate the enzyme solution with double-stranded DNA substrate at
*7℃ for *6h. DNA electrophoresis was used to detect no change in
the double-stranded DNA substrate.
|
reagent
|
usage
amount
|
|
primer
|
X ul
|
|
Oligo(dT)*0
|
final
concentration2.5uM
|
|
Or random
primers
|
final
concentration2.5ng/ul
|
|
Or gene-specific
primers
|
final
concentration0.*5uM
|
|
Template
RNAa
|
*0 ng~1ug/*0ul
|
|
5× M-MLV First Strand Buffer
|
4ul
|
|
0.1 M DTT
|
1ul
|
|
M-MLV GIII Reverse Transcriptase (**0 U/ul)
|
1ul
|
|
dNTP Mix (*0 mM Each)
|
1ul
|
