A simple and rapid modified--new method for SNP typing by fragment length discrepant allele specific PCR / 法医学杂志
Journal of Forensic Medicine
;
(6): 11-14, 2005.
Article
in Chinese
| WPRIM
| ID: wpr-983062
ABSTRACT
OBJECTIVE@#To establish a new method for single nucleotide polymorphism (SNP) typing based on allele specific PCR fragment length discrepant allele specific PCR (FLDAS-PCR), and study the influence on specific extension by introducing a mismatch at the third or fourth 3'-terminal base of allele specific primers.@*METHODS@#For SNP loci rs759117 and rs760887, two allele specific forward primers, with different length and a mismatch introduced at the third or fourth 3'-terminal base, and a public reverse primer were designed for SNP typing. The genotyping of SNP was determined by the two allele specific fragments different in size after polyacrylamide gel and silver staining.@*RESULTS@#The different homozygote genotypes comprised a single band with different size respectively, and the heterozygote genotypes comprised two bands. Typing results were completely consistent with those by direct sequencing. Non-specific primer extension was decreased remarkably after introducing a mismatch at the third or fourth 3'-terminal base of allele specific primers, and the stringency of PCR reaction was cut down.@*CONCLUSION@#FLDAS-PCR is a simple, rapid and efficient new method for SNP typing. During FLDAS-PCR, specific primers with a mismatch at the third or fourth 3'-terminal base have more power to identify two alleles.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Polymorphism, Restriction Fragment Length
/
DNA
/
Polymerase Chain Reaction
/
Sensitivity and Specificity
/
Sequence Analysis, DNA
/
DNA Primers
/
Base Pair Mismatch
/
Polymorphism, Single Nucleotide
/
Electrophoresis, Polyacrylamide Gel
/
Alleles
Limits:
Humans
Language:
Chinese
Journal:
Journal of Forensic Medicine
Year:
2005
Type:
Article
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