Construction of a cDNA library from microdissected guinea pig crista ampullaris

Hear Res. 1994 Feb;73(1):65-6. doi: 10.1016/0378-5955(94)90283-6.

Abstract

Poly(A) RNA was isolated from microdissected guinea pig crista ampullaris epithelium and converted into cDNA with RNase H- murine leukemia virus reverse transcriptase. After size fractionation, the cDNA was directionally ligated into the vector pSPORT 1 and the plasmids electroporated into E. coli. The library was found to have 1.6 x 10(7) independent colonies with 5% of the colonies lacking an insert. Thirty randomly selected colonies were checked for inserts and the average insert size was 833 base pairs with a range of 400 to 2300 base pairs. The library was screened with a beta-actin guinea pig cDNA probe and 0.16% of the colonies contained an insert hybridizing to the probe.

MeSH terms

  • Actins / genetics
  • Animals
  • Base Composition
  • Cloning, Molecular
  • DNA Probes
  • DNA, Complementary / genetics*
  • Gene Library*
  • Guinea Pigs
  • Nucleic Acid Hybridization
  • Semicircular Canals / chemistry*

Substances

  • Actins
  • DNA Probes
  • DNA, Complementary