An acid precipitation technique: A strip assay for the large-scale DNA polymerase activity screening |
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| Authors: | Maria Yu Kordyukova Valeriya N. Antipova Vadim V. Rogachevsky Nadezhda V. Zyrina |
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| Affiliation: | 1. Institute of Molecular Genetics, Russian Academy of Sciences, 123182 Moscow, Russian Federation;2. Institute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russian Federation;3. Institute of Cell Biophysics, Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russian Federation |
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| 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. |
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| Keywords: | DNA polymerase activity screening GF/C glass fiber Bst DNA polymerase Large fragment purification |
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