Journal article
Construction and Preliminary Characterization of a Series of Mouse and Rat Testis cDNA Libraries
Journal of andrology, Vol.20(5), pp.635-639
09/1999
Handle:
https://hdl.handle.net/2376/112106
PMID: 10520576
Abstract
We have constructed a series of 23 cDNA libraries from mouse and rat testicular cells. These include libraries made from whole, intact adult testes; seminiferous tubule cells from adult testes; combined populations of primary spermatocytes from 18‐day‐old mouse testes; and isolated populations of primitive type A spermatogonia, type A spermatogonia, type B spermatogonia, preleptotene spermatocytes, leptotene plus zygotene spermatocytes, juvenile pachytene spermatocytes, adult pachytene spermatocytes, round spermatids, Sertoli cells from 6‐, 8‐, 17‐, and 18–20‐day‐old mice, and peritubular cells from 18–20 day old mice, all recovered from outbred white Swiss (CD‐1) mice. We also constructed libraries from whole adult testes from five other lines of mice: C57 BI6/J, C3 HEB, BDF‐1, Balb/c, and 129 Sv. Finally, there are two libraries made from populations of Sertoli cells and peritubular cells isolated from testes of 20‐day‐old Sprague‐Dawley rats. Enzymatic dissociation, followed by gradient separation or plating/lysing techniques, was used to prepare populations of specific cell types in purities of 85–98%. cDNAs were synthesized from poly A+ mRNA primed with oligo dT and unidirectionally cloned into the lambda Uni‐Zap XR expression vector from Stratagene. Primary titers ranged from 2.1 ± 105 to 2.9 × 108 plaque‐forming units, and insert sizes averaged 1.0–1.2 kb. These libraries have been amplified once and submitted to the American Type Culture Collection (ATCC) for distribution to interested investigators. ATCC accession numbers are provided.
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Details
- Title
- Construction and Preliminary Characterization of a Series of Mouse and Rat Testis cDNA Libraries
- Creators
- JOHN R MCCARREYDEBORAH A o'BRIENMICHAEL K SKINNER
- Publication Details
- Journal of andrology, Vol.20(5), pp.635-639
- Academic Unit
- Biological Sciences, School of
- Publisher
- Blackwell Publishing Ltd; Oxford, UK
- Number of pages
- 5
- Identifiers
- 99900547598001842
- Language
- English
- Resource Type
- Journal article