Abstract
Cellulose-bound DNA complementary to ovalbumin mRNA was used in a continuous hybridization system to isolate single-stranded DNA molecules containing the ovalbumin gene. Fragmented DNA segments containing the ovalbumin gene were enriched 300–350 fold in one cycle of purification. Two cycles of purification resulted in a DNA fraction which was enriched 2300 fold in the ovalbumin sequence. The method is suitable for purification of the ovalbumin sequence from both sheared DNA fragments, as well as larger molecular weight DNA containing more than twice the number of nucleotides necessary to code for ovalbumin mRNA. The chromatographic procedures were specific and reproducible. In addition, the recovery of ovalbumin DNA was essentially quantitative (80–100%), even when large amounts of starting DNA (70–75 mg) were used. This purification scheme should also be useful for the enrichment of other unique sequence genes from eucaryotic DNA.
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