Comparison of different methods of recovering DNA from a methylation assay.
Several enzymes, for example DNA (cytosine-5-)-methyltransferase, produce relatively strong interactions with DNA and hinder the quantitative recovery of this DNA from a reaction mixture. Classical methods like the Sevag chloroform-iso-amylic alcohol one or the phenol procedure lead only to a 50-60 per cent recovery of the DNA. A new procedure was worked out utilizing pancreatic RNase, proteinase K, NaOH 0.5 M treatment and trichloracetic acid precipitation which gives 85 per cent recovery of DNA.
D004247:DNA / Q000302:isolation & purification* / Q000378:metabolism; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism*; D004720:Endonucleases; D019286:Endopeptidase K; D010450:Endopeptidases; D008745:Methylation; D008780:Methyltransferases / Q000378:metabolism*; D019800:Phenol; D010636:Phenols; D012259:Ribonuclease, Pancreatic; D012260:Ribonucleases
D010636:Phenols; D019800:Phenol; D004247:DNA; D008780:Methyltransferases; D004248:DNA (Cytosine-5-)-Methyltransferases; D004720:Endonucleases; D012260:Ribonucleases; D012259:Ribonuclease, Pancreatic; D010450:Endopeptidases; D019286:Endopeptidase K