BTEP maintains several Question and Answer Forums of interest to the NCI/CCR community.
Currently, there are forums on these topics listed below:
If you wish to ask a question go to the Ask Question Page and submit your question.
Library ChIP preparation: What should be the starting amount of DNA for fresh and fixed sample?
What should be the starting amount of DNA for fresh and fixed sample?
1 Answer:
This will really depend on a few empirically derived factors (antibody binding affinity for example). In general, you want to start with a much DNA as possible for each sample whether it is fresh or fixed. The NEBNext ChIP-seq Library Prep Kit recommends 1ng-10ng minimum AFTER immunoprecipitation. Citation: https://www.neb.com/-/media/nebus/files/manuals/manuale6200.pdf?rev=699002accd044e1d80c4977e24df6a07&hash=A25617D7EE043541AA39BD6413A4484F - answered by Tovah Markowitz, Paul Schaughency, Vishal Koparde.
Answered on June 5th, 2020 by amy.stonelake@nih.gov