CONSTRUCTION OF A CDNA LIBRARY FROM MICRODISSECTED GUINEA-PIG CRISTA-AMPULLARIS

Poly(A) RNA was isolated from microdissected guinea pig crista ampulla ris epithelium and converted into cDNA with RNase H- murine leukemia v irus reverse transcriptase. After size fractionation, the cDNA was dir ectionally ligated into the vector pSPORT 1 and the plasmids electropo rated into E. coli. The library was found to have 1.6 x 10(7) independ ent colonies with 5% of the colonies lacking an insert. Thirty randoml y selected colonies were checked for inserts and the average insert si ze was 833 base pairs with a range of 400 to 2300 base pairs. The libr ary was screened with a beta-actin guinea pig cDNA probe and 0.16% of the colonies contained an insert hybridizing to the probe: