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. 2017 Apr 17;26(13):2515–2525. doi: 10.1093/hmg/ddx146

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© The Author 2017. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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In vitro DNA binding and polymerization properties of POLγA:Y955C and POLγA:Y955H. (A) Electrophoretic mobility shift assays showing that POLγA:Y955C and POLγA:Y955H bind DNA both in absence (lanes 5 and 8) and presence (lanes 6 and 9) of POLγB. Lanes 1, 4 and 7 contained no protein. Lower panel: schematic representation of the DNA template. The asterisk indicates the ³²P label on the 5´ end of the 20-mer. (B) Coupled 3’–5’ exonuclease/polymerase assays show that both POLγA:Y955C and POLγA:Y955H required higher dNTP concentrations than WT POLγA to synthesize DNA (all experiments were performed in the presence of POLγB). Reactions were run on denaturing 15% PAGE. Below: scheme of the primer-extension reaction starting from the 20-meric primer to produce a 35-mer product.