Normalization of a cDNA Library Cloned in λZAP by a Long PCR and cDNA Reassociation Procedure
Author(s) -
Katja Fischer,
Deborah C. Holt,
Peter J. Wilson,
James A. Davis,
V. Hewitt,
M. Johnson,
Annette McGrath,
Bart J. Currie,
Shelley F. Walton,
Darrell J. Kemp
Publication year - 2003
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/03342bm03
Subject(s) - library science , cdna library , complementary dna , biology , genetics , computer science , gene
250 BioTechniques Vol. 34, No. 2 (2003) be expected from an amplificationbased strategy. To our knowledge, this is the first time that such analysis has been conducted to assess the representation of a genomic library. In summary, we have developed a quick, facile protocol for constructing libraries from relatively small amounts of DNA. Using our method, we have shown that a representative, random library can be constructed in a single day. It has previously been shown that it is possible to vaccinate an animal using an entire genomic expression library and elicit a protective response (1). Therefore, it may be possible to create a genetic vaccine for even an unknown pathogen in a single day using this method. Given that in certain situations, such as biothreats or outbreaks of zoonoses, there may not be time to identify and isolate protective antigens from a complete genome, immunization with a shotgun random library may be a viable course of action.
Accelerating Research
Robert Robinson Avenue,
Oxford Science Park, Oxford
OX4 4GP, United Kingdom
Address
John Eccles HouseRobert Robinson Avenue,
Oxford Science Park, Oxford
OX4 4GP, United Kingdom