5×LabScript RT MasterMix
This product adopts a new generation of reverse transcriptase with multiple point mutations using molecular evolution technology, greatly improving stability and reverse transcription efficiency. five × RT MasterMix is a one tube reverse transcription premixed reaction solution that contains all the reagents required for reverse transcription (LabScript M-MuLV RTase, RNase Inhibitor, Random primers, Oligo dT Primer, dNTP Mixture, Buffer). The reaction can be carried out by simply adding template RNA and water. This makes the synthesis of cDNA more convenient and efficient, especially suitable for two-step Real Time PCR detection after cDNA synthesis.
First strand cDNA synthesis.
It can be used for the detection of high copy and low copy genes.
The new generation of reverse transcriptases has significantly improved stability and reverse transcription efficiency.
Fully premixed reverse transcription Mix, simply add RNA and water to complete reverse transcription in 15 minutes.
The volume of RNA templates can be added up to 80% of the total volume, making them very suitable for reverse transcription reactions with low concentration RNA templates.
Pre mixed Mix does not freeze at -20 ℃, reducing thawing and mixing time, making it easier to use.
This product specifically optimizes the oligo dT and N6 random primer ratio for qPCR, allowing cDNA synthesis to start from various regions of the RNA transcript and have the same reverse transcription efficiency, ensuring the authenticity and repeatability of qPCR results to the greatest extent possible.
Component 20 μ L × 50 times 20 μ L × 100 times
five × RT MasterMix 200 μ L 400 μ L
RNase free H2O 1mL 2 × 1mL
Storage: -20 ℃, valid for 12 months.
Avoid RNase contamination.
To ensure successful reverse transcription, it is recommended to use high-quality RNA samples.
five × RT MasterMix is very viscous, and the solution can easily adsorb on the tube wall and outside the suction head, causing loss. Before use, please use it after centrifugation and avoid loss of adhesion on the outer wall of the suction head. five × The enzymes contained in RT MasterMix are all excessive, even if 3.6 is used each time μ L-3.8 μ The use of L does not affect the effectiveness of use.
First strand cDNA synthesis:
With 20 μ Taking the L reaction system as an example, 10 can also be used μ L reaction system.
Thaw the template RNA on ice; RNase free H2O is thawed at room temperature (15-25 ℃) and quickly placed on ice after thawing. Before use, replace each
Mix the solution with light bounce or slight vortex oscillation, and briefly centrifuge to collect the remaining liquid on the pipe wall to the bottom of the pipe.
Add the following ingredients to the RNAse free tube: (It is recommended to use a PCR tube to prepare on ice and use a PCR instrument for reaction)
Ingredient dosage
Total RNA/mRNA ≤ 16 μ L
five × RT MasterMix 4 μ L (see note 3)
RNase free H2O to 20 μ L
