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Nucleic Acids Research, 1980, Vol. 8, No. 8 1731-1743
© 1980

Articles

A short primer for sequencing DNA cloned in the single-stranded phage vector M13mp2

S. Anderson, M.J. Gait, L. Mayol* and I.G. Young{dagger}

MRC Laboratory of Molecular Biology Hills Road, Cambridge CB2 2QH, UK

Received February 21, 1980.

In this paper we describe the synthesis and cloning of a short segment of DNA complementary to the region immediately adjacent to the EcoRI insertion site in the single-stranded bacteriophage vector M13mp2. This segment is useful as a "universal" primer for DNA sequencing by the dideoxynucleotide chain termination method; the template can be any DNA species cloned in M13mp2 or its derivatives. The primer has been cloned into the tetracycline resistance gene of plasmid pBR322 as one strand of a 26 bp EcoRI/BamHI fragment. This fragment may be readily prepared from an EcoRI + BamHI restriction digest of the parent plasmid (designated pSPi4) by a simple size fractionation.

*Present address: Instituto di Chimica Organica e Biologica, Universita di Napoli, Napoli, Italy

{dagger}Permanent address: Department of Biochemistry, John Curtin School of Medical Research, Australian National University, Canberra, Australia


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