Process:
Microsatellites are short sequences of nucleotide of 2 to 7 bp tandem repeats, also known as SSR/STR/SSTR, etc. Microsatellites are versatile genetic markers used for studies of ecology, evolution and conservation., etc. The process includes:
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Assay primer design based on gene sequence from data base.
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PCR amplification of desired locus using the labeled primers with or without multiplexing.
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High-throughput fragment analysis of labeled amplicon using ABI 3730xl genetic analyzer system with known standard.
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Data capturing and analysis using Gene Mapper software. Generation of the report containing microsat locus ID, length of product, no. of repeat units and chromatograms, etc.
Sample Requirement
(a)
Purified labeled PCR product (ready to run): 20μl of 10-20ng/μl per sample in 96-well plate, allele size information, level of multiplexing, dye used for multiplexing, amplicon size information (Dyes-FAM, VIC, PET, NED and LIZ(Standard)).
(b)
DNA: 20μl of 150-200 ng/μl of gDNA with quality report, primer information for amplification with standardized conditions for each primer, sizing information, multiplexing details.
Turn Around Time:
10-15 days from the receipt of sample to Xcelris premises.
Deliverables:
Raw data consist of chromatogram file of sizing standard (LIZ-120 or 500), standard curve and allele sizing value. Report consists of allelic information of homozygous and heterozygous status of sample with peak area and height value.
