Product: miRNA Tailing First Strand Synthesis Kit
CAT.No.:MF0202
Price : $ 238 for 20T(negotiable if bulk purchase)
Storage conditions: -20℃, 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 | 20T | 40T |
| RNase free ddH2O | 400μl | 800μl |
| Poly A(yellow) | 20μl | 40μl |
| 2xPoly A Buffer | 200μl | 400μl |
| RTase Mix (red) | 40μl | 80μl |
| 2x RT Buffer | 200μl | 400μl |
Instructions:
1. RNA plus A tail:
Take an RNase-free centrifuge tube and prepare the following mixture:
| RNase free ddH2O | to 20μl |
| Poly A | 1μl |
| 2xPoly A Buffer | 10μl |
| RNA | 10pg~2ug |
Mix gently, incubate at 37℃ for 30min, and quickly place on ice.
2. Preparation of RT reaction solution:
Centrifuge the above reaction solution at 5000rpm for 5s before use:
| Take the reaction liquid | 8μl |
| 2x RT Buffer | 10μl |
| RTase Mix | 2μl |
Use a pipette to mix gently.
3. RT reaction:
| 25℃ | 5min |
| 50℃ | 15min |
| 85℃ | 5min |
After the reaction is completed, centrifuge at 5000rpm 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 -20°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 55°C.
Quality Control
Use the synthesized U6 as a sample, use this kit, add A, reverse transcription, take 1/10 cDNA product for qPCR, and the CT value is less than 20.
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 10pg 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.
