Analysis of MIR200B Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-1340-LGZ)

Introduction

Official Full Name: microRNA 200b
Also known as: MIRN200B; mir-200b
microRNAs (miRNAs) are short (20-24nt) non-coding RNAs that participate in the post-transcriptional regulation of gene expression in multicellular organisms by affecting mRNA stability and translation. miRNAs are transcribed by RNA polymerase II as part of capped and polyadenylated primary transcripts (pri-miRNAs), which can be protein-coding or non-coding. The primary transcript is cleaved by Drosha RNase III to generate an approximately 70 nt stem-loop precursor miRNA (pre-miRNA), which is further cleaved by cytoplasmic Dicer RNase to generate mature miRNA and antisense miRNA star (miRNA*) products. Mature miRNAs are incorporated into the RNA-induced silencing complex (RISC), which recognizes target mRNAs through imperfect base-pairing with miRNAs, most commonly resulting in translational repression or destabilization of target mRNAs. RefSeq represents predicted microRNA stem-loops.

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Principle Applications Procedure Materials Notes Related Products

Principle

Quantitative reverse transcription PCR (RT-qPCR) is an experimental method applied to PCR experiments using RNA as the starting material. In this method, total or messenger RNA (mRNA) is first transcribed into complementary DNA (cDNA) by reverse transcriptase. Subsequently, qPCR reaction was performed using cDNA as template.

Applications

Gene mutation analysis.

Procedure

1. Sample processing and preparation of PCR reaction system.
2. Add the amplification template, cover the PCR reaction cover, mix well, centrifuge at low speed instantaneously, and transfer to the PCR instrument.
3. Set the program for PCR amplification.
4. Data analysis.

Materials

Sample: depends on the customer's analysis requirements

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