A novel strategy for defining haplotypes by selective depletion using restriction enzymes.
Various single nucleotide polymorphisms (SNPs) have been investigated regarding association with gene expression levels or human diseases. Although different SNPs within one gene are frequently analyzed individually, it is highly probable that in the majority of the cases, a precise combination of SNP alleles, i.e., haplotype, determines a functional trait. Methods commonly used for haplotype determination, involving studies in families, cloning, or somatic cell hybrids, are expensive and time-consuming. We herein suggest a novel and simple strategy for haplotype determination, involving selective haplotype depletion with a restriction enzyme, followed by sequencing. We studied 11 LTA gene polymorphisms in 102 Brazilian individuals, and we applied this novel methodology for haplotyping 67 out of 70 LTA heterozygous individuals. We concluded that the method is rapid and efficient, and, as it includes only simple and widespread-used techniques, it could be used in most of the laboratories without further investment in equipments. The wider usage of haplotyping could be important to clarify contradictory results frequently observed among studies that focus on a single SNP.
Smirnova
Anna S
AS
Immunogenetics Division, Pediatrics Department, Universidade Federal de São Paulo (UNIFESP-EPM), Rua Loefgreen, 1235, 04040-031 São Paulo, SP, Brazil. annsmile141@yahoo.com
Ferreira-Silva
Kátia C
KC
Mine
Karina L
KL
Andrade-Oliveira
Vinicius
V
Shulzhenko
Natalia
N
Gerbase-DeLima
Maria
M
Morgun
Andrey
A
D000483:Alleles; D001938:Brazil; D004262:DNA Restriction Enzymes / Q000737:chemistry; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D006239:Haplotypes / Q000235:genetics*; D006801:Humans; D008233:Lymphotoxin-alpha / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide
D008233:Lymphotoxin-alpha; D004262:DNA Restriction Enzymes