Cloning method by multiple-digestion, vectors for...

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

Reexamination Certificate

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C435S006120, C435S091100, C435S091400, C536S025300

Reexamination Certificate

active

06924112

ABSTRACT:
The present invention pertains to a process for isolating an intact clone of one target nucleic acid fragment having a known characteristic, from a group of fragments by preparing an initial library of clones from the group of fragments using a vector containing no more than a predetermined number of known restriction sites, preferably 1–3 restriction sites, subjecting the initial library to at least 10, and preferably between 50 and 70 restriction enzymes different from those to which the vector is susceptible, to produce a group of monodigested libraries, screening the group of monodigested libraries for the target fragment to determine those restriction enzymes to which the target fragment is insensitive, and subjecting the initial library to substantially all of the restriction enzymes to which the target fragment is insensitive, to produce a multidigested library having an intact clone of the target nucleic acid fragment. The target fragment can then be separated, transfected, reproduced, and studied or sequenced.

REFERENCES:
patent: 5252724 (1993-10-01), Kishimoto et al.
patent: WO 95/04745 (1995-02-01), None
Piug Deng and Jac A. Nickoloff, Site-Directed Mutagenesis Of Virtually Any Plasmid By Eliminating A Unique Site.
Jones, et al, Production Of A Vector To FAcilitate DNA Mutagenesis and Recombination, Biotech, vol. 16, No. 4, 1994.
Amersham Life Science Catalogue, pp. 164-165 XP002076172.

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