Suitable spin column for Size selection of RNA for making enriched cDNA library.

Deepak Khandka deepak at BGUMAIL.BGU.AC.IL
Mon Nov 22 10:59:03 EST 1999


Hi,

I want to make a cDNA library to fish out the full length (I hope) cDNA of a approximately 2.5 Kb transcript. My probe is a 1.5 Kb uncharecterized (I have the sequence, but various gene prediction programs gives variable results) genomic DNA fragment. Since I know the size of the transcript (from Northern) I want to make size selection of mRNA (to enrich it with 2.5 Kb range). Is there a suitable spin column in the market for me (lets say that selects RNA > 2 Kb) ? Is it advisable to make the size selection from the agarose gel (in MOPS buffer with formaldehyde) ? Do you have a suitable protocol ?

Also, I am considering to try Clontech's 'SMART III cDNA library construction Kit'. Any advice ?

Thank you.

Deepak Khadka
Ben Gurion University
Sede Boker, Israel 






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