Other Species Collections
pT7T3-PAC Library Method Overview
Libraries in the pT7T3-PAC vector were constructed in the laboratory of Marcello
Bento Soares and M. Fatima Bonaldo, University of Iowa. Briefly, the method was as follows:
- First strand cDNA synthesis was primed with oligo-dT primer containing a Not I site.
- Double stranded cDNA was ligated with an EcoR I adaptor, digested with Not I and then cloned
directionally into the vector.
- Each clone in a given library had a different tag sequence
located between the Not I site and the polyA
tail to identify the library of origin of the clone.
Bonaldo MF, Lennon G, Soares MB. Normalization and subtraction: two approaches to facilitate gene discovery. Genome Research 6:791-806, 1996.