• First strand synthesis kit of cDNA corresponding to miRNA
  • First strand synthesis kit of cDNA corresponding to miRNA

First strand synthesis kit of cDNA corresponding to miRNA

. This kit uses a specially optimized pre mixed miRNA RT Enzyme Mix to combine Poly (A) tailing and reverse transcription into one step, simplifying the operation steps and improving the efficiency of Poly (A) tailing and reverse transcription
$4.20
  • First strand synthesis kit of cDNA corresponding to miRNA

SPECIFICATION

                   First strand synthesis kit of cDNA corresponding to miRNA
. This kit uses a specially optimized pre mixed miRNA RT Enzyme Mix to combine Poly (A) tailing and reverse transcription into one step, simplifying the operation steps and improving the efficiency of Poly (A) tailing and reverse transcription. The kit has the ability to extract from 20pg-2 μμ The first strand of cDNA corresponding to miRNA was effectively prepared from the total RNA of g. A single synthesized cDNA can detect multiple microRNAs, saving samples and costs.
Component specifications
MiRNA RT Enzyme Mix 50 μ L
two × MiRT Reaction Mix 250 μ L
RNase free H2O 1mL
Storage: -20 ℃
1、 MiRNA 3 'terminal undergoes Poly (A) tail addition and reverse transcription reaction (first strand synthesis)
Thawing 2 × MiRNA RT Reaction Mix and mix well. Place miRNA RT Enzyme Mix in ice for later use, and add the following reagents to a total volume of 20 μ L (miRNA RT Enzyme Mix added last).
Component dosage and final concentration
Total RNA x μ L to 2 μ G
two × MiRNA RT Reaction Mix 10 μ L 1 ×
MiRNA RT Enzyme Mix 2 μ L-
RNase free H2O to 20 μ L-
Attention:
① MiRNA RT Enzyme Mix is very viscous, and the solution is prone to adsorption on the tube wall and outside the suction head, resulting in loss. Before use, please gently shake and centrifuge to avoid loss of adhesion on the outer wall of the suction head. Enzymes in Enzyme Mix are always excessive, even if they are treated with 1.8 μ The use of L does not affect the effectiveness of use.
② The total RNA used in the reaction must contain small molecule RNA (miRNA). This process can also use enriched miRNAs. Pure miRNAs cannot be directly quantified using a spectrophotometer, and it is recommended to directly add 2 μ L-5 μ L. The addition amount can be determined based on the abundance of the target miRNA, but for low abundance miRNA samples such as serum plasma extracts, a maximum volume of 8 can be directly added μ L.
Gently mix the prepared reaction solution with a pipette, centrifuge briefly, and react at 42 ℃ for 60 minutes.
Deactivate miRNA RT Enzyme Mix by heating at 85 ℃ for 5 seconds.
The synthesized cDNA reaction solution can be stored at -20 ℃ or directly tested for downstream PCR or fluorescence quantitative PCR.
Note: The cDNA template obtained by following the above steps is used for downstream P
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