High yield of restriction fragment length polymorphisms in two-dimensional separations of human genomic DNA.
We have investigated the extent to which restriction fragment length polymorphism can be detected by two-dimensional electrophoresis of end-labeled genomic restriction fragments. Genomic DNA was digested with NotI and EcoRV and labeled at the NotI recognition site before first-dimension electrophoresis in disk gels. DNA in the disk gels was further digested in situ with HinfI prior to second-dimension electrophoresis, yielding patterns in which approximately 2000 end-labeled fragments were simultaneously visualized. On the basis of studies of 6 mother/father/child trios, a group of 184 fragments was organized into 85 polymorphic systems in which all allelic fragments were detectable in the 2-D patterns. Another 206 fragments varied as to their presence among individuals, but their relatedness to other fragments was not established. Our data indicate that a large number of DNA polymorphisms can be simultaneously scored in 2-D separations of genomic DNA fragments.
Kuick
R
R
Department of Pediatrics, University of Michigan Medical School, Ann Arbor 48109, USA.
Asakawa
J
J
Neel
J V
JV
Satoh
C
C
Hanash
S M
SM
D000328:Adult; D000483:Alleles; D001483:Base Sequence; D002461:Cell Line, Transformed; D002648:Child; D004247:DNA / Q000032:analysis* / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D005787:Gene Frequency; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008297:Male; D008969:Molecular Sequence Data; D010375:Pedigree; D012150:Polymorphism, Restriction Fragment Length
D004247:DNA; D015252:Deoxyribonucleases, Type II Site-Specific; C032229:GANTC-specific type II deoxyribonucleases; C027440:GATATC-specific type II deoxyribonucleases; C038565:GCGGCCGC-specific type II deoxyribonucleases