ChIP-sequencing was performed as previously described 6 (link). 1x107 cells were crosslinked in 1% formaldehyde for 10 minutes and sonicated with a Bioruptor to obtain chromatin fragments of 200–300 bp. IP was performed overnight with the specific antibodies and Dynabeads Protein A or Protein G beads (Life Technologies). Beads were washed and chromatin fragments were eluted in TE with 1% SDS at 65°C. After de-crosslinking overnight, DNA was extracted using Wizard SV extraction columns (Promega) and Illumina sequencing libraries were prepared using NEBNext ChIP-seq library per reagent set (New England Biolabs) and following manufacturer’s instructions. Libraries were assayed on a BioAnalyzer (High Sensitivity DNA kit) and sequenced on a Nextseq500 (Illumina).
Wizard sv extraction columns
Manufactured by Promega
The Wizard SV Extraction Columns are a laboratory equipment product designed for the rapid and efficient purification of DNA from a variety of sample types. These columns utilize a silica-based membrane technology to selectively bind DNA, allowing for the removal of impurities and contaminants. The purified DNA can then be eluted from the column for further downstream applications.
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2 protocols using wizard sv extraction columns
1
ChIP-Seq Protocol for Chromatin Profiling
2
ChIP-Seq Protocol for Chromatin Profiling
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ChIP-sequencing was performed as previously described 6 (link). 1x107 cells were crosslinked in 1% formaldehyde for 10 minutes and sonicated with a Bioruptor to obtain chromatin fragments of 200–300 bp. IP was performed overnight with the specific antibodies and Dynabeads Protein A or Protein G beads (Life Technologies). Beads were washed and chromatin fragments were eluted in TE with 1% SDS at 65°C. After de-crosslinking overnight, DNA was extracted using Wizard SV extraction columns (Promega) and Illumina sequencing libraries were prepared using NEBNext ChIP-seq library per reagent set (New England Biolabs) and following manufacturer’s instructions. Libraries were assayed on a BioAnalyzer (High Sensitivity DNA kit) and sequenced on a Nextseq500 (Illumina).
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