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Conditions Affecting the Isolation from Transformed Cells of Bacillus subtilis of High-Molecular-Weight Single-Stranded Deoxyribonucleic Acid of Donor Origin

¹ Department of Microbiology, The Public Health Research Institute of the City of New York, Inc., New York, New York 10016

ABSTRACT

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.