Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

DNA purification

WI William Jordan, III
EL Erica Larschan
request Request a Protocol
ask Ask a question
Favorite

The samples were cooled to room temperature and 400ul of an equal volume of phenol/chloroform/isoamyl alcohol was added to each sample and the solution was mixed vigorously. The samples were centrifuged for 5 min at 13,000 RPM and the upper aqueous phase was transferred to a new tube. DNA was precipitated by adding 40ul of NaAc pH 5.2 and 1 mL of 100% ethanol. The samples were incubated for 30 min at – 80 °C and then centrifuged at 13,000 RPM for 30 min at 4C. The supernatant was discarded and pellet was washed with 1 mL of ethanol 70%. The sample was centrifuged for 15 min at 13,000 RPM at 4C. The supernatant was discarded and the pellet was air-dried for 5 min. The pellet was resuspended in 1X TE buffer.

DNA shearing and size selection, biotin pull-down and sequencing library preparation were performed as described in [36]. Multiplexed libraries were sequenced on an Illumina Hi-Seq 2500 configured for 150-bp paired-end reads.

Copyright and License information: The Author(s) ©2021
Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

0/150

tip Tips for asking effective questions

+ Description

Write a detailed description. Include all information that will help others answer your question including experimental processes, conditions, and relevant images.

post Post a Question
0 Q&A