Carbon-Copy Chromosome Conformation Capture (5C)
The 5C technique expands from 3C and allows for the parallel analysis of interactions between many selected loci.
Steps 1-4: Same as in 3C.
Step 5 Ligation-mediated amplification and Quantitation: Performing multiplex ligation-mediated amplification (LMA) after the construction of the 3C library leads requires using multiplex primers that consist of universal primer sequences like T7 and T3 and the ligation junction sequences. They anneal to the 3C fragments and get ligated together with a DNA ligase. Perfect alignment with the 3C template ensures the success of the ligation. The ligated primers serve as templates of which get amplified to generate the 5C library. The use of universal primer sequences mean these 5C fragments can be analyzed on microarrays. The small size (~100 bp) of the 5C fragments is also compatible for analysis using high-throughput sequencing.
Read more about this topic: Chromosome Conformation Capture
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