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
Famous quotes containing the words chromosome and/or capture:
“We become male automatically because of the Y chromosome and the little magic peanut, but if we are to become men we need the help of other men—we need our fathers to model for us and then to anoint us, we need our buddies to share the coming-of-age rituals with us and to let us join the team of men, and we need myths of heroes to inspire us and to show us the way.”
—Frank Pittman (20th century)
“Because the young child feels with such intensity, he experiences sorrows that seem inconsolable and losses that feel unbearable. A precious toy gets broken or a good-bye cannot be endured. When this happens, words like “sad” or “disappointed” seem a travesty because they cannot possibly capture the enormity of the child’s loss. He needs a loving adult presence to support him in his pain but he does not want to be talked out of it.”
—Alicia F. Lieberman (20th century)