A Drosophila Complementary DNA Resource-Reference-Cited by-同舟云学术

A Drosophila Complementary DNA Resource

Published:2000-03-24 Issue:5461 Volume:287 Page:2222-2224
ISSN:0036-8075
Container-title:Science
language:en
Short-container-title:Science

Author:

Rubin Gerald M.¹²³,Hong Ling¹³,Brokstein Peter¹³,Evans-Holm Martha¹³,Frise Erwin¹³,Stapleton Mark⁴,Harvey Damon A.¹²³

Affiliation:

1. Berkeley Drosophila Genome Project,

2. Howard Hughes Medical Institute, and

3. Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720–3200, USA.

4. Berkeley Drosophila Genome Project, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.

Abstract

Collections of nonredundant, full-length complementary DNA (cDNA) clones for each of the model organisms and humans will be important resources for studies of gene structure and function. We describe a general strategy for producing such collections and its implementation, which so far has generated a set of cDNAs corresponding to over 40% of the genes in the fruit fly Drosophila melanogaster .

Publisher

American Association for the Advancement of Science (AAAS)

Subject

Multidisciplinary

Reference15 articles.

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2. Total RNA was prepared by hot phenol extraction. mRNA was purified with the Stratagene Poly(A) Quik mRNA isolation kit. mRNA quality was assessed by RNA blots with Delta and Notch clones as probes. First strand cDNA synthesis was carried out with the Stratagene λZAPII-cDNA synthesis kit with the substitution of SuperscriptII reverse transcriptase from GIBCO-BRL. The cDNAs were directionally cloned into the λZAPII and pOT2a vectors. Phage and plasmid libraries were amplified once. The plasmid libraries were size fractionated with GIBCO-BRL S-500 cDNA size fractionation columns and clones with inserts larger than 1 kb were pooled and transformed.

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4. Complementary DNA Sequencing: Expressed Sequence Tags and Human Genome Project

5. 5′ ESTs were generated with either dye primer or dye terminator chemistries on ABI373 and ABI377 sequencers.

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