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PCR Topics pp 53-55 | Cite as

The Use of Short Adapters for Priming PCR of Unknown Chromosomal DNA Fragments

  • J. Schneider
  • H. Schrempf

Abstract

For the analysis of unknown stretches of deletable DNA from Streptomyces lividans 1326 [3,4], we plan to apply the following protocol for substractive hybridization [1]: randomly sheared substracter DNA from the variant strain Nl (with the respective DNA deleted) is biotinylated [5], mercurated [2] and hybridised in great surplus with unlabelled wild-type DNA fragments generated by a Sau3A partial digest. Subsequent affinity chromatography on a Thio-propyl-Sepharose 6B (Pharmacia) and Streptavidin-Agarose (Gibco-BRL) double column removes all DNA hybrids containing at least one strand of labelled DNA, leaving a solution with a small amount of wild-type specific fragments, each only in a few copies. Thus, prior to further studies, these fragments have to be amplified considerably.

Keywords

Streptavidin Agarose Double Column Preliminary Positive Result Short Adapter Pharmacia Column 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

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Copyright information

© Springer-Verlag Berlin Heidelberg 1991

Authors and Affiliations

  • J. Schneider
  • H. Schrempf

There are no affiliations available

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