Screening of SSR Primers by Capillary Electrophoresis (CAT#: STEM-ET-0304-ZJF)

Introduction

Simple sequence repeats (SSRs) are ubiquitous short tandem duplications found within eukaryotic genomes. Their length variability and abundance throughout the genome has led them to be widely used as molecular markers for crop-breeding programs, facilitating the use of marker-assisted selection as well as estimation of genetic population structure. The number of available SSR primers in various plant species varies from a few to several hundred.
In this service, a small number of samples were selected to screen several pairs of microsatellite primers. First, the M13 joint primer sequence was synthesized to test the primers, and primers with high efficiency and stable amplification and polymorphism were selected. Then, fluorescently labeled primers were synthesized, and a mixed-detection scheme of primer combinations would be designed for genetic diversity analysis.

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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, SSR-primer

Procedure

1. The primer sequence is provided to synthesize M13 connector primers
2. Genome extraction
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.
Soil: Dry or wet, dry soil effect is better, quality > 5g
Paraffin section: > 10 slices of 10*10*8 um, transported at room temperature.

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