Selector-technique
Encyclopedia
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 or RCA.
DNA
Deoxyribonucleic acid is a nucleic acid that contains the genetic instructions used in the development and functioning of all known living organisms . The DNA segments that carry this genetic information are called genes, but other DNA sequences have structural purposes, or are involved in...
is digested with restriction enzymes, circularized by hybridisation to selectors and subsequently attached to a vector sequence by ligation
Ligation
Ligation may refer to:* In molecular biology, the covalent linking of two ends of DNA molecules using DNA ligase* In medicine, the making of a ligature * Chemical ligation, the production of peptides from amino acids...
. The procedure results in circular DNA molecules with an included general primer pair motif that can be used for amplification by PCR or RCA.
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.Selection mechanisms
- (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 polymeraseTaq polymerasethumb|228px|right|Structure of Taq DNA polymerase bound to a DNA octamerTaq polymerase is a thermostable DNA polymerase named after the thermophilic bacterium Thermus aquaticus from which it was originally isolated by Thomas D. Brock in 1965...
.