RT Journal Article
SR Electronic
T1 Quantification of DNA samples by Ethidium Bromide Spot Technique
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 289108
DO 10.1101/289108
A1 Jessica Zhao
A1 Julian Zhao
A1 Antoine Cummins
A1 Tiffany Gonzalez
A1 Grace Axler-DiPerte
A1 Devin Camenares
YR 2018
UL http://biorxiv.org/content/early/2018/03/27/289108.abstract
AB Accurate and quick determination of DNA concentration is critical for the assembly of synthetic constructs, as well as a multitude of other experiments. We sought to optimize an under-utilized and inexpensive approach for determining DNA concentration: a spotting technique that uses the intercalating dye Ethidium Bromide. This technique does not require specialized equipment such as a spectrophotometer, but instead relies on visualization of dye-DNA complex fluorescence when excited by UV light. We modelled and tested a range of parameters for dye concentration and spot size, finding that 15uL spots with 1.0ug/mL Ethidium Bromide produced the most reliable standard curve. More importantly, we hope that our approach can help other labs optimize this protocol for their own experimental setup. Adoption of this technique may help enable development of iGEM teams in resource limited environments and laboratories which do not or cannot employ a satisfactory method for determining DNA concentration.Financial Disclosure We obtained funding support from Canon Solutions America, and indirect support for this work through the CUNY Research Scholars Program. However, the funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Competing Interests The authors have declared that no competing interests exist.Ethics Statement N/AData Availability Yes – all data are fully available without restriction. The data can be found in the associated supplementary materials.Article Tags DNA, Ethidium Bromide, EtBr, Concentration, Method, UV, Gibson AssemblyHeader Statement This work was assessed during the iGEM/PLOS Realtime Peer Review Jamboree on 23rd February 2018 and has been revised in response to the reviewers. Responses to Reviewers, and supplementary information (including Fig. S1) are available as separate files.