QuestionsCategory: ChIP-Seq Data AnalysisLibrary ChIP preparation: What should be the starting amount of DNA for fresh and fixed sample?
stonelakeak@nih.gov Staff asked 5 months ago

What should be the starting amount of DNA for fresh and fixed sample?

1 Answers
stonelakeak@nih.gov Staff answered 5 months ago

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.