Direct haplotype-specific DNA sequencing.
Prep Biochem Biotechnol
; 36(3): 253-7, 2006.
Article
en En
| MEDLINE
| ID: mdl-16707336
ABSTRACT
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.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Haplotipos
/
ADN
/
Análisis de Secuencia de ADN
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
Prep Biochem Biotechnol
Asunto de la revista:
BIOQUIMICA
/
BIOTECNOLOGIA
Año:
2006
Tipo del documento:
Article
País de afiliación:
Noruega