Analysis of Mutant CEP290 mRNAs by RT-qPCR (CAT#: STEM-MT-0058-LGZ)

Introduction

Leber Congenital Amaurosis (LCA) is a severe inherited retinal dystrophy responsible for congenital or early-onset blindness. The most common pathogenic mutation (>10%) is located deep in intron 26 of the CEP290 gene (c.2991+1655A>G). It creates a strong splice donor site, leading to the insertion of a cryptographic exon encoding a premature stop codon.

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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

Leber Congenital Amaurosis (LCA)

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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