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Purification of nanogram-range immunoprecipitated DNA in ChIP-seq application.

著者 Zhong J , Ye Z , Lenz SW , Clark CR , Bharucha A , Farrugia G , Robertson KD , Zhang Z , Ordog T , Lee JH
BMC Genomics.2017 Dec 21 ; 18(1):985.
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Chromatin immunoprecipitation-sequencing (ChIP-seq) is a widely used epigenetic approach for investigating genome-wide protein-DNA interactions in cells and tissues. The approach has been relatively well established but several key steps still require further improvement. As a part of the procedure, immnoprecipitated DNA must undergo purification and library preparation for subsequent high-throughput sequencing. Current ChIP protocols typically yield nanogram quantities of immunoprecipitated DNA mainly depending on the target of interest and starting chromatin input amount. However, little information exists on the performance of reagents used for the purification of such minute amounts of immunoprecipitated DNA in ChIP elution buffer and their effects on ChIP-seq data. Here, we compared DNA recovery, library preparation efficiency, and ChIP-seq results obtained with several commercial DNA purification reagents applied to 1 ng ChIP DNA and also investigated the impact of conditions under which ChIP DNA is stored.
PMID: 29268714 [PubMed - in process]
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