Detection of DNA Microsatellite Instability (MSI) by Fluorescent PCR-Capillary Electrophoresis (CAT#: STEM-ET-0303-ZJF)

Introduction

A microsatellite is a tract of repetitive DNA in which certain DNA motifs (ranging in length from one to six or more base pairs) are repeated, typically 5-50 times. Microsatellites are often referred to as short tandem repeats (STRs) or as simple sequence repeats (SSRs). Microsatellite instability (MSI) is the condition of genetic hypermutability (predisposition to mutation) that results from impaired DNA mismatch repair (MMR). It is closely related to the occurrence of tumor.
In this service, MSI is detected by fluorescent PCR-capillary electrophoresis. DNA from normal tissue and tumor tissue was extracted at the same time, and the detection sites were amplified by multiple fluorescent PCR technology, and then the amplified products were detected by capillary electrophoresis. Finally, the detection results of the two tissue sources would be compared and analyzed by professional software.

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Principle Applications Procedure Materials Notes Related Products

Principle

Capillary Electrophoresis (CE) is physical method of analysis which performs in a separation channel of elastic quartz capillary, under the influence of a high voltage direct current field. Charged analytes dissolved in an electrolyte solution are separated based on differences in mobility and/or distribution behavior of components. The migration velocity of an analyte under an electric field is determined by the electrophoretic mobility of the analyte and the electro-osmotic mobility of the buffer inside the capillary. The electrophoretic mobility of a solute depends on the characteristics of the solute (electric charge, molecular size and shape) and those of the buffer in which the migration takes place (type and ionic strength of the electrolyte, pH, viscosity and additives). Capillary electrophoresis provides greater resolution, higher sensitivity and online detection. It enables single-cell analysis and even single-molecule analysis, optimizing separation and analysis of biological macromolecules.

Applications

Genomics, clinical analysis, tumor, cancer

Procedure

1. DNA extraction and purification
2. DNA template quantification
3. PCR amplification
4. Capillary electrophoresis
5. Data analysis

Materials

• Capillary electrophoresis apparatus
• Sample solution
• Buffer solution

Notes

Sample requirements:
Genomic DNA: Concentration > 20-30ng/ul, OD260/280 about 1.7-2.1, transported by ice pack.
Animal tissue: > 500mg, transported at low temperature.
Blood (anticoagulant): > 1ml, transported at low temperature.
Cell: > 10^6, collect cell precipitate, dry ice transport.
Paraffin section: > 10 slices of 10*10*8 um, transported at room temperature.

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