miRNA cDNA 第一链合成预混液（茎环法）

miRNA cDNA first strand synthesis premix (stem-loop method)

Product Code: AG11748

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¥700

Product Description

This is a reverse transcription kit for miRNA 1st Strand cDNA synthesis by stem-loop method. This product is optimized for the reaction system. 5X miRNA 1st Strand cDNA Synthesis Premix (Stem-loop) contains the buffer and the buffer for the reverse transcription reaction. M-MLVIt is easy to use and can effectively reduce the loss of reagents and experimental errors, and shorten the operation time by adding only the stem-loop primer, RNA template and water. This product is suitable for reverse transcription of miRNA samples such as Total RNA or Small RNA. The cDNA synthesized by this product is suitable for chimeric qPCR analysis and is recommended to be used with SYBR Green. Pro Taq HS Premixed qPCR Kit II (Code No. AG11702); for high demand of primer dimer elimination, it can be used with SYBR Green Pro Taq HS Premixed qPCR Kit IV (Code No. AG11746) for superior results.

Product Advantages

①This product is a premixed product, the reaction solution is very simple to prepare, only need to add the stem-loop primer, RNA template and water can be directly carried out the reaction.
②This product can reverse transcribe samples containing miRNA such as Total RNA or Small RNA.
③ The cDNA obtained by inversion of this product can realize accurate quantification of mature miRNA with high sensitivity and specificity.

Product Composition
individual parts making up a compound norm
5X miRNA 1st Strand cDNA Synthesis Premix (Stem-loop) 200 μl
RNase free water 1 ml

 

Preservation and transportation
Storage temperature: -20℃ storage
Transportation temperature:Dry ice transportation or -20°C ice pack transportation.
Example 1

Mouse liver Total RNA (1 μg, 100 ng, 10 ng, 1 ng, 100 pg, 10 pg, 0 pg, respectively) was used as a template for reverse transcription experiments using this product, and 2 μl of cDNA stock solution was taken from the cDNA obtained by reversing the cDNA as a template, which was then analyzed by SYBR Green. Pro Taq HS Premixed qPCR Kit II (Code No. AG11702 ) was used to perform qPCR amplification for the detection of mice. mmu-miR-29a-3p。
Quantitative instrument used: ABI QuantStudioTM 5 Real-Time PCR Systems. the results are shown below:

The results are shown above:
1、Working curve R2= 0.9992 and the amplification efficiency was 92.61 TP3T.
2. The amplification curves in the range of 1 μg ~ 10 pg RNA concentration showed good linearity.
3、Single melting curve peak type, good amplification specificity.

Example 2

Mouse liver Total RNA (1 μg, 100 ng, 10 ng, 1 ng, 100 pg, 10 pg, 1 pg, 0 pg, respectively) was used as a template for reverse transcription experiments using this product, and 2 μl of the cDNA stock solution was taken as a template for cDNA obtained by reverse transcription, and then the cDNA stock solution was extracted with SYBR Green. Pro Taq HS Premix qPCR Kit II (Code No. AG11702) was used to detect qPCR amplification in mice. tRF RNA-mmu_tsr000557。
Quantitative instrument used: ABI QuantStudioTM 5 Real-Time PCR Systems. the results are shown below:

The results are shown above:
1、Working curve R2= 0.9985 and the amplification efficiency was 103.91 TP3T.
2. The amplification curves in the range of 1 μg ~ 1 pg RNA concentration showed good linearity.
3、Single melting curve peak type, good amplification specificity.

Example 3

293T Total RNA (1 μg, 100 ng, 10 ng, 1 ng, 100 pg, 10 pg, 0 pg, respectively) was used as a template for reverse transcription experiments using this product, and 2 μl of cDNA stock solution was taken from 2 μl of cDNA obtained by reversing the cDNA as a template and analyzed by SYBR Green. Pro Taq The HS Premix qPCR Kit II (Code No. AG11702) was used for qPCR amplification to detect human miR-221-3p。
Quantitative instrument used: ABI QuantStudioTM 5 Real-Time PCR Systems. the results are shown below:

The results are shown above:
1、Working curve R2= 0.9991 and the amplification efficiency was 92.71 TP3T.
2. The amplification curves in the range of 1 μg ~ 10 pg RNA concentration showed good linearity.
3、Single melting curve peak type, good amplification specificity.

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