Subtractive PCR?
Robert Brown
rcbrown at casbah.acns.nwu.edu
Wed Dec 18 17:44:47 EST 1991
In article <1991Dec18.213347.27747 at magnus.acs.ohio-state.edu> cbeall at magnus.acs.ohio-state.edu (Clifford J Beall) writes:
>Dear Methods people,
>
>Does anyone have a method for subtracting cDNA prior to PCR?
>We are using degenerate primers and want to clone a cell specific
>message. It seems one could hybridize with [+] cDNA with [-] RNA,
>remove hybrids, then treat the remainder with RNAse. Is HAP still the
>method of choice for hybrid removal and does anyone know conditions
>for this? Thanks in advance.
> Cliff Beall
> Ohio State Biotech. Ctr.
Invitrogen sells a kit for subtractive hybridization using biotinylation of
the [-] RNA and extraction of the hybrid into phenol after binding
streptavidin.
good luck
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