Selector technique: Difference between revisions

Revision as of 09:41, 4 October 2017

Revision as of 09:45, 4 October 2017

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{{Unreferenced|date=October 2017}}{{Orphan|date=February 2009}}

The '''selector technique''' is a method to [[DNA amplification|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.

= Process =

==The selector 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 [[PCR]] or [[Rolling circle replication|RCA]].

==The selectorSelector construct==

A selector consists of two oligonucleotides, one '''Vector''' oligonucleotide and one '''Selector probe'''. Together they form one '''Selector''' with target specific ends on each side of a general primer motif.

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==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==