Genomic analysis II: isolation of high molecular weight heteroduplex DNA following differential methylase protection and Formamide-PERT hybridization.
Understanding the nature of DNA sequence differences among individuals is important to the understanding of fundamental questions in biology. To analyze such differences in complex genomes new approaches must be developed. We report two new techniques which aid in this effort. First, we have developed a modification of the Phenol Emulsion Reassociation Technique (PERT) that allows hybridization of long (20 kb and longer) single copy heteroduplex DNA fragments from human genomic DNAs. Secondly, by using a differential methylase protection technique we have shown that double methylase resistant heteroduplex DNA molecules can be size fractionated away from reannealed single methylase resistant homoduplex DNA molecules. These methods will be useful in obtaining DNA from chromosomal subregions linked to the inheritance of a specific trait or condition as described in the preceding paper and could also be used to create a map of the chromosomal subregion which includes the gene for the trait.
D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism*; D004262:DNA Restriction Enzymes; D005796:Genes; D006801:Humans; D008970:Molecular Weight; D009691:Nucleic Acid Denaturation; D009692:Nucleic Acid Heteroduplexes / Q000235:genetics* / Q000302:isolation & purification; D009693:Nucleic Acid Hybridization; D019800:Phenol; D010636:Phenols
D009692:Nucleic Acid Heteroduplexes; D010636:Phenols; D019800:Phenol; D004247:DNA; D004248:DNA (Cytosine-5-)-Methyltransferases; D004262:DNA Restriction Enzymes