Construction of a cDNA library from microdissected guinea pig crista ampullaris.
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.
Wilcox
E R
ER
Laboratory of Molecular Biology, National Institute on Deafness and Other Communication Disorders, Bethesda, MD 20892.
Fex
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J
D000199:Actins / Q000235:genetics; D000818:Animals; D001482:Base Composition; D003001:Cloning, Molecular; D015342:DNA Probes; D018076:DNA, Complementary / Q000235:genetics*; D015723:Gene Library; D006168:Guinea Pigs; D009693:Nucleic Acid Hybridization; D012665:Semicircular Canals / Q000737:chemistry*
D000199:Actins; D015342:DNA Probes; D018076:DNA, Complementary