Conditions affecting the isolation from transformed cells of Bacillus subtilis of high-molecular-weight single-stranded deoxyribonucleic acid of donor origin.
The deoxyribonucleic acid (DNA) extraction procedure of Piechowska and Fox was evaluated to determine which steps are required for the isolation of high-molecular-weight single-stranded material from transformed cultures of Bacillus subtilis. The results indicate that high-molecular-weight single-stranded DNA can be isolated when certain basic proteins are present at the time of lysis. In the absence of such protective agents as lysozyme or cytC the single-stranded DNA is degraded. The single-stranded DNA can also be protected by being treated with lysozyme at low temperature. The high molecular weight of this single-stranded material and its kinetics of appearance are consistent with its being an intermediate in the transformation process.
D001412:Bacillus subtilis / Q000032:analysis*; D002250:Carbon Radioisotopes; D002499:Centrifugation, Density Gradient; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification*; D004277:DNA, Single-Stranded / Q000032:analysis / Q000302:isolation & purification*; D008970:Molecular Weight; D009113:Muramidase; D009691:Nucleic Acid Denaturation; D013696:Temperature; D013936:Thymidine; D014170:Transformation, Genetic; D014316:Tritium
D002250:Carbon Radioisotopes; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D014316:Tritium; D009113:Muramidase; D013936:Thymidine