Section 5
Chapter 4,574

A family of lambda phage complementary dna cloning vectors lambda swaj allowing the amplification of rna

Palazzolo, M.J.; Meyerowitz, E.M.

Gene 52(2-3): 197-206


ISSN/ISBN: 0378-1119
Accession: 004573123

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This paper describes the construction and characterization of a family of .lambda. 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 hybrdization and in situ hybridization procedures. Using substrative hybridization and this vector system, 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.

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