Fluorscein- Fluorimager how to - (Jul/03/2006 )
I use Chemicon TRAPeze XL for telomerase detection. If the assay is positive, we should get a ladder of DNA fragments each with 6 bp difference. The advantage of this is that after PCR amplification the product can be visualised directly by Fluooimaging system as the amplifluor primers are labelled with Fluorescein. Am not sure how to go about imaging. plz suggest.
My immediate thought is to find someone with a Phosphorimager you can use.
Most of them have a blue laser (450 nm) which can be used to excite and detect fluorescein (488 nm). We use a Storm 840 but the Storm 860 and Typhoon classes will do this as well. You may find that the gel (agarose or acrylamide?) is too thick to get a really decent image. In which case you need to dry it on to a non-fluorescent surface.