Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1997 Nov 1;25(21):4427–4428. doi: 10.1093/nar/25.21.4427

A method for generating subtractive cDNA libraries retaining clones containing repetitive elements.

C T Lin 1, D R Sargan 1
PMCID: PMC147059  PMID: 9336480

Abstract

Here we describe a two-stepped photobiotin-based procedure to enrich a target (canine retinal) cDNA library for tissue specific clones without removing those containing repetitive ( SINE ) elements, despite the presence of these elements in the driver population. In a first hybridization excess SINE elements were hybridized to a driver (canine cerebellar) cDNA. In a second hybridization target cDNA was added to this reaction. The resulting cDNA library was enriched for retinal specific clones, but contained the same ratio of clones with SINE elements found in the unsubtracted library.

Full Text

The Full Text of this article is available as a PDF (38.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  2. Duguid J. R., Rohwer R. G., Seed B. Isolation of cDNAs of scrapie-modulated RNAs by subtractive hybridization of a cDNA library. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5738–5742. doi: 10.1073/pnas.85.15.5738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Minnick M. F., Stillwell L. C., Heineman J. M., Stiegler G. L. A highly repetitive DNA sequence possibly unique to canids. Gene. 1992 Jan 15;110(2):235–238. doi: 10.1016/0378-1119(92)90654-8. [DOI] [PubMed] [Google Scholar]
  4. Schweinfest C. W., Henderson K. W., Gu J. R., Kottaridis S. D., Besbeas S., Panotopoulou E., Papas T. S. Subtraction hybridization cDNA libraries from colon carcinoma and hepatic cancer. Genet Anal Tech Appl. 1990 May;7(3):64–70. doi: 10.1016/0735-0651(90)90042-e. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES