Sizing of single‐stranded regions in double‐stranded DNA by preparative benzoylated DEAE‐cellulose chromatography

FEBS Letters

Norris, Murray D.¹  Stewart, Bernard W.¹ 

[1] Children's Leukaemia and Cancer Research Unit, Prince of Wales Children's Hospital, Randwick, NSW 2031, Australia

关键词: DNA structure;    Benzoylated DEAE-cellulose;    Single-stranded region;    Chromatography;    Nucleotide length;    DNA hybrid;    BND-cellulose;    benzoylated naphthoylated DEAE-cellulose;    BD-cellulose;    benzoylated DEAE-cellulose;   

DOI  :  10.1016/0014-5793(88)80003-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The concentration of caffeine required to elute wholly single-stranded DNA from benzoylated DEAE-cellulose is proportional to the polynucleotide length. The use of benzoylated DEAE-cellulose chromatography for isolating and sizing single-stranded regions in double-stranded DNA has been examined using a series of hybrid molecules. Restriction fragments of the replicating form of bacteriophage ϕX174 were hybridized to the intact ‘plus’ strand, thereby forming hybrids having single- and/or double-stranded regions in the kilobase range. A series of such hybrid preparations were subject to caffeine concentration gradient elution from benzoylated DEAE-cellulose. After logarithmic transformation, a linear relationship (R = 0.94) could be demonstrated between eluting caffeine concentration and single-stranded length, irrespective of the length of associated double-stranded regions or the location, within a given fragment, of unpaired nucleotides. Benzoylated DEAE-cellulose chromatography may therefore be used to separate and characterize, on a preparative scale, double-stranded DNA containing single-stranded regions.

【 授权许可】

Unknown