Selector technique: Difference between revisions

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The '''selector technique''' is a method to [[polymerase chain reaction|amplify]] and [[Multiplexing|multiplex]] genomic DNA.

The '''selector technique''' allows multiplex amplification of arbitrary sets of genomic sequences. Genomic [[DNA]] is digested with restriction enzymes, circularized by [[hybridisation (molecular biology)|hybridisation]] to selectors and subsequently attached to a vector sequence by [[DNA ligase|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.

==The selectorProcess construct==

The '''selector technique''' allows multiplex amplification of arbitrary sets of genomic sequences. Genomic [[DNA]] is digested with [[restriction enzymes]], circularized by [[hybridisation (molecular biology)|hybridisation]] to selectors and subsequently attached to a [[Plasmid|vector]] sequence by [[DNA ligase|ligation]]. The procedure results in circular DNA [[molecules]] with an included general [[Primer (molecular biology)|primer]] pair motif that can be used for amplification by [[Polymerase chain reaction|PCR]] or [[Rolling circle replication|RCA]].

==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.

A selector consists of two oligonucleotides, one '''Vector''' oligonucletideoligonucleotide 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 polymerase]].

==Publications==

*[https://web.archive.org/web/20051127101236/http://nar.oxfordjournals.org/cgi/content/full/33/8/e71 Demonstration of the selector method]

*[https://archive.today/20130415163620/http://nar.oxfordjournals.org/cgi/content/full/33/8/e72 The PieceMaker software for designing selector experiments]

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