Magnetic RNA isolation
anmab2 at orion.alaska.edu
Sun Mar 3 03:15:06 EST 1996
BMB6MWS at leeds.ac.uk ("M.W. SMITH") wrote:
>Can anybody comment on their experiences with mRNA isolation using
>magnetic beads e.g. from Promega or Dynal. I need to make high quality
>poly a+ RNA for construction of a cDNA library. These manufacturers claim
>that their kits can be used for direct isolation of poly A+ from tissue,
>without the need to prepare total RNA. Does this provide high quality
>poly a+? Any comments would be gratefully received.
>Mark Smith (m.w.smith at leeds.ac.uk)
>University of Leeds.
I've used the Promega PolyAttract 1000 kit for isolating polyA(+) RNA directly from whole Xenopus embryos with good results in both 5' RACE and cDNA library construction. The one CRITICAL thing I know of with the magnetic particle technology is that the magnetic ferritin/oligo dT particles must not be frozen in shipment or they will not work at all. Depending on where you are (I'm in Alaska, so you can imagine), that may be a consideration. The only problem that I've observed with them other than that of freezing is trace contamination of the RNA with magnetic particles after isolation, so an additional spin to pellet out the remaining particles is probably a good idea, though they supposedly don't cause problems in small quantities in most procedures.
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