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  • Title: In vitro production of large single-stranded templates for DNA sequencing.
    Author: Straus NA, Zagursky RJ.
    Journal: Biotechniques; 1991 Mar; 10(3):376-84. PubMed ID: 1648367.
    Abstract:
    A method has been developed for the preparation of large single-stranded DNA sequencing templates from primary cloning plasmids or cosmids. The method utilizes the separate action of T7 Gene 6 exonuclease and exonuclease III to generate large quantities of single-stranded template for each strand of a large-cloned fragment. Since the procedure is intended for use on primary clones, it avoids the time-consuming subcloning steps associated with most sequencing programs. The procedure also has the advantage of avoiding clone instability problems associated with subcloning in M13.
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