Hi--
I need references and help from people familiar with alternative
splicing. I've isolated alternatively spliced cDNAs from a MHC class I
gene which appear fairly "normal," i.e., they are spliced exon to exon.
However, I've also isolated a fair number of cDNAs from the same prep
that are spliced in a rather haphazard fashion, e.g., the middle of one
exon is spliced to the 3'UTR, others have deletions near intron/exon
boundaries.
As our lab has little experience in studying splicing, I want to know if
these are "real," or are they splicing intermediates, or could they be
experimental artifact (these were all subcloned from RT/PCR reactions
from total RNA)? Would we see any difference if we isolate message
only? Does anyone have an excellent reference on the subject?
Thanks,
jon