The value of supplement of recombinational Tth-single strand binding protein in polymerase chain reaction for genotyping / 中华检验医学杂志
Chinese Journal of Laboratory Medicine
;
(12): 233-236, 2012.
Artigo
em Chinês
| WPRIM
| ID: wpr-428562
ABSTRACT
ObjectiveTo explore the value of Tth-single strand binding protein (SSB) used as an additive to improve the polymerase chain reaction (PCR) specificity for single nucleotide polymorphisms(SNP) alleles genotyping.MethodsTth-SSB plasmid was constructed and the protein was expressed,then the expressed Tth-SSB was added into PCR system detecting cytochrome P450 Protein ( CYP2C19 * 3,636G>A)genotype to determine the optimal usage and condition.Then,the genotypes of 30 cerebral ischemia patients were tested with established methods and compared with direct sequencing to verify the accuracy of Tth-SSB as an additive into PCR for SNP genotyping.ResultsThe purity of Tth-SSB was 85% and optimal dosage was 1 μg.The protein could improve the specificity and reduce the dimer when Tth-SSB was added into the PCR system.Thirty patients genotyping results as follow26 patients belong to G/G homozygote,4 patients belong to G/A heterozygote,no body belong to A/A homozygote.The coincidence acquire 100% with parallel sequencing.ConclusionAs an additive,Tth-SSB could significantly improve the accuracy of genotyping by eliminating non-specific bands.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
Chinese Journal of Laboratory Medicine
Ano de publicação:
2012
Tipo de documento:
Artigo
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