RT Journal
A1 Houge, G
T1 Simplified construction of a subtracted cDNA library using asymmetric PCR.
JF Genome Research 
JO Genome Research 
YR 1993 
FD February 01 
VO 2 
IS 3 
SP 204 
OP 209 
DO 10.1101/gr.2.3.204 
UL http://genome.cshlp.org/content/2/3/204.abstract 
AB A novel method for the direct construction of subtracted plasmid cDNA libraries in the plasmid pBluescript is presented. Two libraries in lambda-ZAP were compared starting with general phagemid excision from both libraries. Thereafter, single-stranded (ss) plasmids from one library were subtracted with biotinylated cDNA molecules generated by asymmetric PCR on ss plasmid templates from the other library. The nonsubtracted plasmids were used to transform Escherichia coli directly, thus making a subtracted plasmid library. Preliminary data suggest that the specificity of the method is around 25%. The method is sensitive enough to detect low-abundance mRNAs. In contrast to other subtractive methods based on lambda-ZAP, the bias introduced using PCR in this case only affects the method's specificity and not its sensitivity.