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pYX-Asc Library Method Overview

Denatured RNA was size fractionated on a 1% agarose gel. First strand cDNA synthesis was primed with oligo-dT primer containing a Not I site. Double strand cDNA was size selected according to mRNA size fraction, ligated with EcoR I adaptor, digested with NotI and then cloned directionally into the pYX-Asc vector.

Reference:

Bonaldo MF, Lennon G, Soares MB. Normalization and subtraction: two approaches to facilitate gene discovery. Genome Research 6:791-806, 1996.