2 replies to this topic

[chan8]

I am interested in amplifying the heavy and light chain from a hybridoma. I have been attempting to amplify them using RT-PCR, but I was curious as to whether these same primers would work in PCR directly on cells and genomic DNA. After V(D)J rearrangement, I would think that the sequences of the genomic DNA would be identical to the sequence of the cDNA, as all the introns would have already been removed in the rearrangement.

Is my reasoning flawed? Thank you in advance.

[Chelo]

Chan8,

there are still introns in the genome, even though arrangements have taken place.

[chan8]

Chelo, on 16 April 2012 - 10:06 AM, said:

Chan8,

there are still introns in the genome, even though arrangements have taken place.

Thank you for your response Enthusiast. I was under the impression that after a B Cell undergoes VDJ rearrangement, that the gene segments were brought together, and that the introns were therefore removed.

Edited by chan8, 17 April 2012 - 10:17 AM.