proteinz, on Aug 1 2009, 03:28 AM, said:
hi guys! i am having a hair pulling moment here i cant determine what my northern blot results mean i hope someone can help...
i ran c. elegans RNA and detected with cDNA probe. the length of the (cdna) gene is 594bp. my gDNA sequence is approx 900bp . i am getting two transcripts at approx 900bp and 1200bp what does this mean?
thanks in advance
what are their relative intensities to one another?
how specific is your probe? do you get 2 signals on a Southern?
both could be your gene....size determination on denaturing gels is approximate, however, since the cDNA is predicted to be ~ 600bp you cannot be sure about the modifications your transcript might be subjected to.
the sizes could be due to variable length poly-A tail, variable 5' upstream elements, closely related gene, multiple copy gene, non-specific target, poor probe design, overloaded total RNA causing the ribosomal bands to appear on your film/phosphorimager etc.