PCR based targeted genomic and cDNA differential display

We previously described a targeted genomic differential display method (TGD D: Broude NE, Chandra A, Smith CL. Differential display of genomic subsets containing specific interspersed repeats. Proc. Natl. Acad. Sci. USA 1997,9 4:4548-53). In that method, presently characterized as method I, targeting was accomplished bq capturing DNA fragments containing specific a sequence by hybridization with complementary single-stranded DNA. The captured fragm ents were amplified by PCR. Here, we describe method II where targeting is accomplished by PCR using primers specific to the target sequence. Method I I rakes advantage of PCR suppression to eliminate fragments not containing the target sequence (Siebert PDA, Chenchik A, Kellogg DE, Lukyanov KA and L ukyanov SA, An improved PCR method for walking in uncloned genomic DNA. Nuc leic Acids Res 1995;23:1087-1088), Targeting fetuses analysis on and around interesting areas and additionally serves to reduce the complexity of the amplified subset. These approaches are useful to amplify genome subsets con taining a variety of targets including various conserved sequences coding f or cis-acting elements or protein motifs. (C) 1999 Published by Elsevier Sc ience B.V. All rights reserved.