Unit

UNIT 6.2 Identifying Transcribed Sequences in Arrayed Bacteriophage or Cosmid Libraries

  1. Ute Hochgeschwender,
  2. Miles B. Brennan

Published Online: 1 MAY 2001

DOI: 10.1002/0471142905.hg0602s00

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Hochgeschwender, U. and Brennan, M. B. 2001. Identifying Transcribed Sequences in Arrayed Bacteriophage or Cosmid Libraries. Current Protocols in Human Genetics. 00:6.2:6.2.1–6.2.15.

Author Information

  1. National Institute of Mental Health, Bethesda, Maryland

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: FEB 1994

Abstract

This unit describes methods for identifying bacteriophage or cosmid clones that contain sequences present in the mRNA of a cell line or tissue. In the Basic Protocol, a radiolabeled cDNA probe is synthesized by reverse transcription of poly(A)+ RNA isolated from the cell line or tissue of interest. The probe is incubated with DNA-cellulose to remove highly repetitive sequences before it is used for hybridization analysis of filters from a phage or cosmid genomic library. After identification of positive clones from the library screen, the same probe can be used to screen Southern blots of restriction enzyme-digested DNA from the positive clones. Support protocols describe preparation and testing of DNA-cellulose.