Published 2006

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Publication details

Journal : Preparative Biochemistry & Biotechnology , vol. 36 , p. 253–257–5 , 2006

International Standard Numbers :
Printed : 1082-6068
Electronic : 1532-2297

Publication type : Academic article

Contributors : Rudi, Heidi; Gylder, Knut-Erik; Rognli, Odd Arne; Rudi, Knut

Issue : 3

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Kjetil Aune
Chief Librarian


Determining haplotype-specific DNA sequence information is very important in a wide range of research fields. However, no simple and robust approaches are currently available for determining haplotype-specific sequence information. We have addressed this problem by developing a very simple and robust haplotype-specific sequencing approach. We utilise the fact that DNA sequencing polymerases are sensitive to 3'end mismatches in the sequencing primer. By using two sequencing primers with 3'end corresponding to the two alleles in a given SNP locus, we are able to obtain allele-specific DNA sequences from both alleles. We evaluated this direct haplotype-specific approach by determining haplotypes within the intron 2 sequence of the fructan-6-fructosyltransferase (6-ft) gene in Lolium perenne L. We obtained reliable haplotype-specific sequences for all primers and genotypes evaluated. We conclude that the haplotype-specific sequencing is robust, and that the approach has a potentially very wide application range for any diploid organism.