Cloning regions of the Drosophila genome by microdissection of polytene chromosome DNA and PCR with nonspecific primer (original) (raw)
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Department of Biology, Princeton University
Princeton, NJ 08544, USA
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Department of Biology, Princeton University
Princeton, NJ 08544, USA
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Division of Oncology, State University of New York
Stony Brook, NY 11794, USA
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Received:
27 September 1989
Revision received:
12 December 1989
Accepted:
12 December 1989
Published:
11 February 1990
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Cedric S. Wesley, Mathew Ben, Martin Kreitman, Nabil Hagag, Walter F. Eanes, Cloning regions of the Drosophila genome by microdissection of polytene chromosome DNA and PCR with nonspecific primer, Nucleic Acids Research, Volume 18, Issue 3, 11 February 1990, Pages 599–603, https://doi.org/10.1093/nar/18.3.599
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Abstract
A simple and rapid procedure to isolate clones carrying sequences from a specific region of the polytene chromosome of Drosophila is demonstrated. The procedure involves microdissection of the region of interest, amplification of the DNA by PCR using a primer designed to prime the synthesis nonspecifically, labeling of the amplified DNA using the random primer method, and screening of a standard library with the probe to identify and isolate clones carrying sequences homologous to the dissected region. This procedure has the potential to replace the difficult procedure of microcloning, as well as facilitate chromosome walking.
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