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Polymerase chain reaction haplotyping using 3′ mismatches in the forward and reverse primers: application to the biallelic polymorphisms of tumor necrosis factor and lymphotoxin
Journal article   Peer reviewed

Polymerase chain reaction haplotyping using 3′ mismatches in the forward and reverse primers: application to the biallelic polymorphisms of tumor necrosis factor and lymphotoxin

G.C. Fanning, M. Bunce, C.M. Black and K.I. Welsh
Tissue Antigens, Vol.50(1), pp.23-31
1997
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Abstract

A polymerase chain reaction with sequence-specific primers (PCR-SSP) system that operates under identical conditions to HLA phototyping was devised for characterizing polymorphisms in tumor necrosis factor (TNF) and lymphotoxin α (LT-α). Mismatches at the 3′ end were incorporated into the forward and reverse primers of each PCR so as to unequivocally establish the cis / trans status between the biallelic sites. Three previously described biallelic polymorphisms in TNF and three in LT-α were characterized in a 24-reaction PCR-SSP system. The method was used to genotype 20 cell lines and 201 HLA class I and II typed controls from the United Kingdom at the TNF and LT-α loci. Population frequencies of TNF haplotypes were determined as was linkage disequilibrium with HLA-A, B, Cw, DRB1 and DQB1 loci. In each gene there were 8 theoretical polymorphic combinations; 4 were observed in TNF and 4 in LT-α. A total of 11 TNF-LT-α haplotypes were determined from apparent homozygous controls and statistical analysis.

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Citation topics
1 Clinical & Life Sciences
1.6 Immunology
1.6.351 Sepsis Immunology
Web Of Science research areas
Cell Biology
Immunology
Pathology
ESI research areas
Molecular Biology & Genetics
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