Which of the Following Experimental Procedures is NOT Used in ChIP-Seq

Question

Which of the following experimental procedures is NOT used in ChIP-seq?
A) DNA-protein complexes are cross-linked.
B) DNA is extracted and sheared.
C) Bead-bound antibodies to transcription factors are used to precipitate transcription factors cross-linked with DNA.
D) The bead-bound antibodies are precipitated using a secondary antibody.
E) Transcription factors are released from DNA, amplified and identified by sequencing.

Accepted Answer

The correct procedure for ChIP-seq involves cross-linking DNA-protein complexes, shearing DNA, using bead-bound antibodies to precipitate transcription factors cross-linked with DNA, and then sequencing the DNA fragments bound to the transcription factors. The use of a secondary antibody to precipitate bead-bound antibodies is not a standard step in ChIP-seq protocols.