A family of lambda phage cDNA cloning vectors, λSWAJ, allowing the amplification of RNA sequences
Abstract
This paper describes the construction and characterization of a family of λ phage cDNA cloning vectors that allows high-efficiency directional cDNA cloning and selective amplification of either sense or antisense cRNA sequences. These vectors contain several unique restriction sites (EcoRI, XbaI, and SacI) positioned between two specific phage promoters, SP6 and T7. This system facilitates the in vitro preparation of single-stranded (ss) RNA molecules that should be useful in subtractive hybridization and in situ hybridization procedures. Using subtractive hybridization and this vector system, it should be possible to identify sequences present in one cDNA library and not another. In addition, it should be possible to construct subtracted cDNA libraries in these vectors and to generate high specific activity, ss, antisense cRNA probes directly from DNA prepared from the whole subtracted library or from individual clones.
Additional Information
© 1987 Elsevier. Received 18 November 1986, Accepted 22 December 1986. We would like to thank James A. Cummer of Central Arizona College for many helpful discussions and members of the E.M.M. laboratory for helpful technical advice. Thanks also to C. Chang for the Arabidopsis alcohol dehydrogenase clone, P. Pang for E. coli GW3810, and to K. Fryxell, C. Chang, P. Mathers, C. Martin, M. Garfinkel, and K. Vijay Raghavan for careful reading of the manuscript prior to publication. This work was supported by NIH program project grant GM20927. M.J.P. was supported by a postdoctoral fellowship from the ...Additional details
- Eprint ID
- 72782
- Resolver ID
- CaltechAUTHORS:20161213-143336780
- GM-20927
- NIH
- Created
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2016-12-13Created from EPrint's datestamp field
- Updated
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2021-11-11Created from EPrint's last_modified field