タイトルまたは説明
Product: miRNA Tailing First Strand Synthesis Kit
CAT.No.:MF***2
Price : $ **8 for *0T(negotiable if bulk purchase)
Storage conditions: **0℃, valid for 1 year, avoid repeated
freezing and thawing
Product Introduction
miRNA PolyA+ RT Kit uses genetically engineered Poly A polymerase,
which can efficiently add Poly A tail to miRNA for subsequent RT
reaction. The reverse transcription primer is a specially designed
primer containing Oligo d(T), which has high affinity for miRNA
containing Poly A tail, so that the RT reaction can be carried out
efficiently. The RT efficiency of miRNA PolyA+ RT Kit (gDNA-) is
better than that of many similar products. In particular, when used
with LABLEAD Bio miRNA PolyA SYBR qPCR Kit, you can get the
matching qPCR downstream primers for free and more ideal
experimental results.
Product Features
Poly A polymerase optimized by genetic engineering, miRNA Poly A
polymerization is efficient and fast; specially designed primers
containing Oligo d(T) can efficiently bind to miRNA Poly A+, and
the RT efficiency is higher than that of many similar reagents;
The same RNA can obtain RT products of multiple miRNAs in one RT,
and multiple miRNAs can be tested later, saving sample usage.
Product Components
|
component
|
*0T
|
*0T
|
|
RNase free
ddH2O
|
**0μl
|
**0μl
|
|
Poly
A(yellow)
|
*0μl
|
*0μl
|
|
2xPoly A
Buffer
|
**0μl
|
**0μl
|
|
RTase Mix
(red)
|
*0μl
|
*0μl
|
|
2x RT
Buffer
|
**0μl
|
**0μl
|
Instructions:
1. RNA plus A tail:
Take an RNase-free centrifuge tube and prepare the following
mixture:
|
RNase free
ddH2O
|
to
*0μl
|
|
Poly
A
|
1μl
|
|
2xPoly A
Buffer
|
*0μl
|
|
RNA
|
*0pg~2ug
|
Mix gently, incubate at *7℃ for *0min, and quickly place
on ice.
2. Preparation of RT reaction solution:
Centrifuge the above reaction solution at ***0rpm for 5s before
use:
|
Take the
reaction liquid
|
8μl
|
|
2x RT
Buffer
|
*0μl
|
|
RTase
Mix
|
2μl
|
Use a pipette to mix gently.
3. RT reaction:
|
*5℃
|
5min
|
|
*0℃
|
*5min
|
|
*5℃
|
5min
|
After the reaction is completed, centrifuge at ***0rpm for
5s, place on ice as soon as possible, and directly perform PCR
reaction. If not used temporarily, please place it at a temperature
below **0°C and use it within half a year. Do not freeze and thaw
repeatedly. If the template has a complex secondary structure or a
high GC region, the reaction temperature can be increased to
*5°C.
Quality Control
Use the synthesized U6 as a sample, use this kit, add A, reverse
transcription, take 1/*0 cDNA product for qPCR, and the CT value is
less than *0.
Notes:
1 The utensils and water used for RT reaction should be RNase
inactivated, and the operator should wear a mask, gloves, etc. to
prevent RNase contamination.
2 Template RNA usage: Total RNA can be between *0pg and 2ug.
3 It is recommended to prepare the reagent on crushed ice. The
viscosity of RTase Mix is
​​relatively high, so it should
be sucked slowly. The Tip head should not be inserted too deep into
the liquid to avoid sticking too much enzyme solution.
