An acid precipitation technique: A strip assay for the large-scale DNA polymerase activity screening

Anal Biochem. 2016 Nov 15:513:39-42. doi: 10.1016/j.ab.2016.08.022. Epub 2016 Aug 26.

Abstract

A new format of a very rapid, low-cost and high-productive analysis based on the acid precipitation of radiolabeled DNA was developed. By contrast to the conventional processing of a large number of GF/C discs, the method employs one GF/C strip containing samples on individual teeth. The strip assay was validated by comparison with the glass fiber disk technique; the efficiency was demonstrated by screening E. coli superproducers and fractions obtained at the steps of Bst DNA polymerase, Large Fragment purification by the protocol we developed. The principle proposed allows simultaneous assaying many samples for the activity of different polymerases.

Keywords: Bst DNA polymerase; DNA polymerase activity screening; GF/C glass fiber; Large fragment purification.

MeSH terms

  • Bacterial Proteins / chemistry*
  • DNA, Bacterial / chemistry*
  • DNA-Directed DNA Polymerase / chemistry*
  • Escherichia coli / chemistry*
  • Geobacillus stearothermophilus / enzymology*

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • DNA-Directed DNA Polymerase