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Analysis of LOC110599582 Gene (Mutation) by RT-qPCR (CAT#: STEM-MT-1585-LGZ)

Introduction

Gene description: CEB1 minisatellite repeat instability region<br />Gene type: biological region<br />This region is a highly variable small satellite region that overlaps with the GAL3ST2 gene on the q arm of chromosome 2. This minisatellite is an unstable GC-rich variable number tandem repeat (VNTR) with a repeat unit length of 39 nucleotides that is similar but not identical. The consensus repeat motif (tcagcccagggacctccgcagcccccccccccccccc) contains the prdm9a binding motif (LD hotspot motif) CCNCCNTNNCCNC. Meiotic recombination hotspots overlapping with repeat regions were found. Instability at this site was observed in germ cells with a strong paternal bias in mutation rates. There is a correlation between allele length and mutation rate, and the allele with more repeats has a higher mutation rate. Both intra-allelic and inter-allelic rearrangements have been observed, with intra-allelic events occurring mainly in homogeneous segments of the repeat region, and inter-allelic events displaying a slight polarity, with more rearrangements occurring at the centromere-proximal end of the minisatellite.




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