PCR & visualise products on gel

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This presentation was originally prepared by C. William Birky, Jr. Department of Ecology and Evolutionary Biology The University of Arizona It may be used.

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Presentation transcript:

PCR & visualise products on gel Figure S12 Chromosome Conformation Capture =3C Live cells are incubated with formaldehyde to crosslink proteins to DNA. cross link Nuclei are purified and exposed to detergent to allow the DNA to be digested with restriction enzymes, e.g. EcoRI digest After digestion the reaction volume is increased to dilute out non-crosslinked DNA. DNA that is in close proximity is expected to be linked by the ligase. A no ligase control ensures that the products are not due to cross-hybridisation with other genomic sequences. ligate reverse cross-link & purify The bait and target primer pairs are used in PCR reactions. The products are visualised on a gel and sequenced. bait target PCR & visualise products on gel