Cloning regions of the Drosophila genome by microdissection of polytene chromosome DNA and PCR with nonspecific primer (original) (raw)

Journal Article

Cedric S. Wesley ,

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Mathew Ben ,

2

Department of Biology, Princeton University

Princeton, NJ 08544, USA

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Martin Kreitman ,

2

Department of Biology, Princeton University

Princeton, NJ 08544, USA

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Nabil Hagag ,

1

Division of Oncology, State University of New York

Stony Brook, NY 11794, USA

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Walter F. Eanes

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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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