The selector technique
allows multiplex amplification of arbitrary sets of genomic sequences. Genomic DNA
is digested with restriction enzymes, circularized by hybridisation
to selectors and subsequently attached to a vector sequence by ligation
. The procedure results in circular DNA molecules
with an included general primer pair motif that can be used for amplification by PCR
The selector construct
A selector consists of two oligonucleotides, one Vector oligonucletide and one Selector probe. Together they form one Selector with target specific ends on each side of a general primer motif.
- (I) A selector probe hybridizes with both ends of the selected target.
- (II) A selector probe hybridizes with one end to the 3’ end of the target and the other end to an internal sequence of the target. The protruding 5' end is cleaved off using Taq polymerase.