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Nucleic Acids Research, 1980, Vol. 8, No. 23 5541-5549
© 1980

MOLECULAR BIOLOGY

A subcloning strategy for DNA sequence analysis

A.M. Frischauf, H. Garoff and H. Lehrach

European Molecular Biology Laboratory Postfach 102209, 6900 Heidelberg, GFR

Received October 20, 1980. We describe here a new strategy of fragment preparation for sequencing procedures using endlabelled DNA fragments as substrates (2,3) which is directly applicable to DNA fragments cloned into the Pst I site of pBR322, or in modified form, to inserts into the BamH I or Sal I site of the same plasmid. Ordered sets of subclones of predetermined overlap are generated. These can be sequenced directly without further strand- or fragment separation steps.


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