|Table of Contents|

A METHOD OF SNP GENOTYPING IN SOYBEAN(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2007年04期
Page:
447-453,459
Research Field:
Publishing date:

Info

Title:
A METHOD OF SNP GENOTYPING IN SOYBEAN
Author(s):
YUAN Cui-ping; LI Ying-hui; LIU Zhang---xiong; GUAN Rong-xia; CHANG Ru-zhen; QIU Li-juan
Chinese Academy of Agricultural Sciences,Institute of Crop Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement,Key Laboratory of Crop Germplasm & Biotechnology (MOA),Beijing 10008.
Keywords:
Soybean; SNP; Genotyping; Fragment Length Discrepant Allele Specific PCR
PACS:
S565.1
DOI:
10.3969/j.issn.1000-9841.2007.04.001
Abstract:
Single nucleotide polymorphism (SNP) is a genome-wide mutation type for organisms.SNP genotyping is an important technique for SNP genetic mapping,association analysis,and molecular assistant selection and so on.We introduced a fragment length discrepant allele specific PCR (FLDAS-PCR) method for SNP genotyping in soybean,using 5 SNPs in Rhg4 as examples.For a SNP loci,we designed 2 allele specific primers which had 4-5bp length difference and 3’-end complementary to each of bi-allelic SNPs,and 1 common primer,as a result PCR products of different alleles had 4-5bp difference in length,which could be separated by 6% denature polyacrylamide gel.In order to improve PCR specificity and data accuracy,an additional base mismatch was introduced at the 3rdor 4thnucleotide closest to the 3’-end of each of the specific primers.We also investigated the influence of annealing temperature and primer concentration on PCR amplification,Rhg4-1592 as an example,and found that lower concentration of the specific primer would reduce its annealing competition in PCR reaction,and behaved more sensitive to annealing temperature,so we could optimize PCR by increasing or reducing specific primer concentration.The genotypes of SNPs by the FLDAS-PCR agreed with the results by PCR cloning and sequencing for 18 soybean genotypes,which approved that the method was effective in SNP genotyping in soybean.

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Last Update: 2014-10-20