Unit

UNIT 6.3 Direct Selection of cDNAs Using Genomic Contigs

  1. Michael Lovett

Published Online: 1 MAY 2001

DOI: 10.1002/0471142905.hg0603s00

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Lovett, M. 2001. Direct Selection of cDNAs Using Genomic Contigs. Current Protocols in Human Genetics. 00:6.3:6.3.1–6.3.15.

Author Information

  1. University of Texas Southwestern Medical Center at Dallas, Dallas, Texas

Publication History

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

Abstract

In the Basic Protocol, uncloned cDNA or cDNA from a library with highly repetitive sequences suppressed are hybridized to biotinylated genomic DNAs. The selected sequences are amplified by PCR and rehybridized to create a secondary-selected enriched cDNA library. Biotinylation of cloned genomic DNA is presented in the first support protocol. Linker addition and suppression of repeats in uncloned DNA is presented in the second support protocol and insert amplification and suppression of repeats from libraries is detailed in the third support protocol.In the Basic Protocol, uncloned cDNA or cDNA from a library with highly repetitive sequences suppressed are hybridized to biot.