High-Stringency Subtraction for the Identification of Differentially Regulated cDNA Clones

The technique of high-stringency subtraction described here facilitates subtractive hybridizations between directional cDNA libraries constructed in λ ZAP® II cloning vectors and represents an improvement on earlier methods for the subtraction of entire cDNA libraries. High-stringency subtraction is...

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Bibliographic Details
Main Authors: Charles P. Scutt, Philip M. Gilmartin
Format: Article
Language:English
Published: Taylor & Francis Group 1997-09-01
Series:BioTechniques
Online Access:https://www.future-science.com/doi/10.2144/97233st04
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Summary:The technique of high-stringency subtraction described here facilitates subtractive hybridizations between directional cDNA libraries constructed in λ ZAP® II cloning vectors and represents an improvement on earlier methods for the subtraction of entire cDNA libraries. High-stringency subtraction is designed to eliminate the subtraction of differentially expressed cDNAs, which show similarity to constitutive sequences by the incorporation of a novel high-stringency wash step. This method also allows the size-selection of target cDNAs and incorporates an improved procedure for the synthesis of driver DNA used in subtractions.
ISSN:0736-6205
1940-9818