Detection and Quantification of Mitochondrial DNA Deletions in Individual Cells by RT-qPCR (CAT#: STEM-MT-0040-LGZ)

Name: Detection and Quantification of Mitochondrial DNA Deletions in Individual Cells by RT-qPCR
SKU: STEM-MT-0040-LGZ
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Introduction

Mitochondrial DNA (mtDNA) defects are an important cause of disease and play a role in the aging process. In a cell, there are multiple copies of the mitochondrial genome. In many patients with acquired or inherited mtDNA mutations, a mixture of mutant and wild-type genomes (known as heteroplasmy) exists within a single cell. As biochemical and clinical defects are only observed when high levels of mtDNA mutations are present, determining heterogeneous mutation levels within tissues and single cells is a critical study.

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Principle Applications Procedure Materials

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

Detection of genetic mutations.

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.