There are many techniques available to enquire about the oganization of the nucleus. I am keenly interested to know the basic machanism behind it. I mean how does it works?
Answers
0
Hi Rahul,
Following are the initial step in 3C and 3C-derived methods to establish a representation of the 3D organization of the DNA.
To this end, the chromatin is fixed using a fixative agent, most often formaldehyde. Next, the fixed chromatin is cut with a restriction enzyme recognizing 6 base pairs (bp)—such as HindIII, BglII, SacI, BamHI, or EcoRI—or with more frequent cutters, such as AciI or DpnII. In the subsequent step, the sticky ends of the cross-linked DNA fragments are religated under diluted conditions to promote intramolecular ligations (i.e., between cross-linked fragments). DNA fragments that are far away on the linear template, but colocalize in space, can, in this way, be ligated to each other. A template is thereby created that is, in effect, a one-dimensional (1D) cast of the 3D nuclear structure. Find more at http://genesdev.cshlp.org/content/26/1/11.full
Hi Rahul,
Following are the initial step in 3C and 3C-derived methods to establish a representation of the 3D organization of the DNA.
To this end, the chromatin is fixed using a fixative agent, most often formaldehyde. Next, the fixed chromatin is cut with a restriction enzyme recognizing 6 base pairs (bp)—such as HindIII, BglII, SacI, BamHI, or EcoRI—or with more frequent cutters, such as AciI or DpnII. In the subsequent step, the sticky ends of the cross-linked DNA fragments are religated under diluted conditions to promote intramolecular ligations (i.e., between cross-linked fragments). DNA fragments that are far away on the linear template, but colocalize in space, can, in this way, be ligated to each other. A template is thereby created that is, in effect, a one-dimensional (1D) cast of the 3D nuclear structure. Find more at http://genesdev.cshlp.org/content/26/1/11.full
Hope useful.
Cheers