Construction of a pulsed-field map around D3S3 using partially demethylated DNA.
Heavy methylation of restriction sites in the relevant chromosomal region can be a major problem in the construction of a long-range restriction map. In the region around the locus D3S3 there appeared to be few accessible restriction sites. Therefore, we cultured cells in the presence of 5-azacytidine and used the partially demethylated DNA to construct a relatively detailed restriction map spanning approximately 1 Mb. Using partially demethylated DNA is a recommendable approach when a genome region appears inaccessible for pulsed-field analysis because excessively long restriction fragments are obtained.
van der Hout
A H
AH
Department of Medical Genetics, University of Groningen, The Netherlands.
Hulsbeek
M M
MM
Buys
C H
CH
D001374:Azacitidine; D002874:Chromosome Mapping / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D003640:Dealkylation; D016521:Electrophoresis, Gel, Pulsed-Field; D006801:Humans
D004247:DNA; D001374:Azacitidine