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miRNA Reagents

miRcute Plus miRNA First-Strand cDNA Kit

Faster and more sensitive reagent for reverse transcription of miRNA by tailing method

Catalog number / packaging

Cat. no

ID

No. of preps

4992909

 KR211-02 20 µl×50 rxn

4992786

 KR211-01 20 µl×25 rxn
Manual Inquire Inspection report

  • Storage

    Store at -20°C

  • Description

    The miRcute Plus miRNA cDNA First-Strand cDNA Kit adopts the A-tailing method to carry out reverse transcription of miRNA first strand cDNA. The kit adopts a two-component form, thus simplifying the experimental operation process and reducing the possibility of operation errors. miRNA RT Enzyme Mix in this product includes E.coli Poly(A) Polymerase, RTase and RNasin. The E. coli Poly (A) Polymerase not only has high A-tailing efficiency, but also can specifically recognize single-stranded miRNA, thus avoiding further reverse transcription reaction of miRNA precursors with double stranded structures. The RTase has been molecular modified to remove RNase H activity and increase the affinity of RNA template, thus enabling miRNA reverse transcription reaction to have higher efficiency and sensitivity. The 2×miRNA RT Reaction Buffer in this product contains all the raw materials and primers for miRNA A-tailing reaction and reverse transcription reaction. It is carefully optimized to ensure that the Poly(A) modification process and reverse transcription process at the 3’ end of miRNA can be efficiently carried out at the same time.

  • Features

    ■ Time-saving and labor-saving: A-tailing reaction and reverse transcription reaction is combined in one step, reducing the operation steps and saving half of the reaction time.

    ■ Strong specificity: Specific modification and reverse transcription of single-stranded miRNA to avoid the influence of miRNA precursor with secondary structure.

    ■ High sensitivity: All miRNA A-tailing products are used for further reverse transcription reaction, greatly improving the detection rate of low abundance miRNA.

    ■ Widely applicable: Reverse transcription reaction can be carried out on miRNA extracted from almost all materials. The input range of templates can reach 20 pg-2 μg (mass range) and 10 fM-100 pM (concentration range).

  • Applications

    ■ Real time quantitative study of miRNA by dye-based/probe-based method

  • Important Notes

    ■ This kit must be used together with miRcute Plus miRNA qPCR Kit (SYBR Green) (4992887/4992779)

Experimental Example

  • Figure. 1. Reverse transcription of human miRNA mimic mix with TIANGEN miRcute Plus miRNA First-Strand cDNA Kit and similar products of Supplier Q (left) Supplier T (right), respectively. The hsa-miR-133a-3p was detected by real time PCR. The results show that for the same RNA sample, the amount of cDNA obtained after reverse transcription of TIANGEN miRcute Plus miRNA First-Strand cDNA Kit (red line) is larger than that of other Suppliers (blue line), indicating higher RT efficiency.

    miRNA mimic mix sample amount: 2×108 copies; Reverse transcription system: 20 μl. 2 μl was sent for quantification; Quantitative reagent: TIANGEN miRcute Plus miRNA qPCR Kit (SYBR Green) (4992887/4992779); Instrument: ABI 7500 fast.

  • Figure 2. Use TIANGEN miRcute Plus miRNA First-Strand cDNA Kit (top row) and relevant product from Supplier Q (lower row) to reverse transcribe human total RNA respectively, and then hsa-miR-122-5p is quantitatively detected by real time PCR. The results show that for the same RNA sample, TIANGEN miRcute Plus miRNA First-Strand cDNA Kit has higher sensitivity and stability than Supplier Q, and is more suitable for downstream experiments.

    Total RNA of Human Huh7 cell is extracted by TRNzol Universal (4992730). Reverse transcription loading: 100ng-1 pg; Reverse transcription system: 20 μl. 2 μl was sent for quantification; Quantitative reagent: TIANGEN TIANGEN miRcute Plus miRNA qPCR Kit (SYBR Green) (4992887/4992779); Instrument: ABI 7500 fast.

Publication

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