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. 2018 Apr 17;5:107. doi: 10.3389/fmed.2018.00107

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Copyright © 2018 Smith, Gajsiewicz and Morrissey.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Traces of polyphosphate (polyP) in cell-derived DNA. DNA isolated from HEK 293 cells using the DNeasy Blood & Tissue kit was extensively digested with benzonase to hydrolyze the DNA, concentrated, and then resolved by electrophoresis on a 4–20% polyacrylamide gel. Samples were: a locally prepared polyP ladder (lengths indicated in phosphate units); DNA purified from HEK 293 cells and digested with benzonase (“Pre”); the same material following digestion with calf intestinal alkaline phosphatase (CIAP); and 50 bp DNA ladder. The same gel was stained sequentially, using: (A) DAPI with extended photobleaching to detect polyP (27) and (B) SYBR Green I to detect DNA (after removing DAPI by repeated rinsing). The material in the lane marked “Pre” is clearly polyP as it photobleached rapidly (A), was digested by CIAP (A), and did not stain with SYBR Green 1 (B).