miRNA Reagents

miRcute Plus miRNA qPCR Kit(SYBR Green)

Higher specificity and sensitivity

Catalog number / packaging

Cat. no

ID

No. of preps

4992779

FP411-02 20 µl×500 rxn

4992887

FP411-01 20 µl×125 rxn
  • Storage

    Store in dark at -20°C
  • Description

    The kit adopts the principle of SYBR® Green I chimeric fluorescence to carry out miRNA fluorescence quantitative detection. The kit contains all reagents for quantitative miRNA fluorescence detection, including 2×miRcute Plus miRNA Premix, 50×ROX Reference Dye and Reverse Primer. Among them, 2× miRcute Plus miRNA Premix (With SYBR & ROX) is a new generation of premixed fluorescent quantitative PCR detection reagent specially developed for miRNA quantitative detection. The DNA Polymerase uses chemically modified hot-start polymerase, cooperating with a special buffer system, it has better reaction specificity, higher sensitivity, and accurate quantification can be carried out in a wider range.
  • Features

    ■ High sensitivity: Target miRNA of total RNA of pg grade can be detected.

    ■ Strong specificity: The completely upgraded system can better distinguish single base differences and perfectly distinguish different miRNA within family.

    ■ Good versatility: Suitable for various types of real time PCR instruments.

  • Applications

    ■ Detect reverse transcribed cDNA of miRNA by real time PCR
  • Important Notes

    ■ This kit must be used together with miRcute Plus miRNA First-Strand cDNA Kit (4992786/4992909).

Experimental Example

  • Table 1. There are 10 kinds of 3p miRNA in the human let-7 family, with little or even single base difference among them. The 10 miRNA mimics were used as templates and different primers were used to amplify the 10 miRNA respectively by using the TIANGEN miRcute Plus miRNA qPCR Kit (SYBR Green). The results show that the specific primer only amplifies the specific template, and the non-specific recognition rate is less than 5%, indicating that TIANGEN miRcute Plus miRNA qPCR Kit (SYBR Green) has good specificity and can distinguish miRNA with single base difference.
  • Figure. 1. Use TIANGEN miRcute Plus miRNA qPCR Kit (SYBR Green) and similar products of Supplier Q, E and T to quantify hsa-miR-15a-5p respectively. The amplification curve, melting curve and standard curve results show that, for the same cDNA sample, TIANGEN miRcute Plus miRNA qPCR Kit (SYBR Green) has higher specificity, sensitivity and linear range, and its quantification is more accurate than that of other suppliers. Total RNA of Human 293T cells was extracted with TRNzol Universal (4992730) and 1 μg was taken for reverse transcription. TIANGEN miRcute Plus miRNA First-Strand cDNA Kit (4992786) was used for reverse transcription, with 20 μl reaction system. After reverse transcription, cDNA was diluted 10-105 times respectively, and 2 μl was taken for quantification. The instrument used is ABI 7500 fast.