NET-Seq

NG Capture-C

NG Capture-C is a refinement of 3C-Seq and Hi-C. It represents a family of methods used to analyze chromatin interactions. NG Capture-C adds multiple pull-down steps of the biotinylated fragments with magnetic beads to the 3C method. The protocol uses formaldehyde fixation followed by restriction enzyme digestion and ligation to form ~10 kb concatamers. The DNA is extracted and sonicated. Indexing adapters are added, and the samples are pooled, purified by pull-down, and PCR-amplified. The pull-down and PCR steps can be repeated to yield up to a million-fold enrichment.

Pros:
  • High sensitivity to detect cis and trans interactions
  • Low sample input requirements
  • Sonicated capture fragments reduces cost compared to capture-C
  • Sonicated capture fragments act as UMIs to reduce PCR bias
Cons:
  • None reported