Selector technique: Difference between revisions

Revision as of 04:54, 20 February 2009

Latest revision as of 00:26, 24 November 2022

(14 intermediate revisions by 13 users not shown)

Line 1:

{{OrphanUnreferenced|date=FebruaryOctober 20092017}}

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 [[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 [[Polymerase chain reaction|PCR]] or [[Rolling circle replication|RCA]].

====The selectorSelector 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====

====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]].

* (I)# A selector probe hybridizes with both ends of the selected target.

====Publications====

* (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====

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

*[https://web.archive.org/web/20051127101236/http://nar.oxfordjournals.org/cgi/content/full/33/8/e72 The PieceMakere71 softwareDemonstration forof designingthe selector experimentsmethod]

*[https://archive.today/20130415163620/http://nar.oxfordjournals.org/cgi/content/full/33/8/e71e72 The PieceMaker Demonstrationsoftware offor thedesigning selector methodexperiments]

{{DEFAULTSORT:Selector-Technique}}

[[Category:DNA]]