Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes
<p>Abstract</p> <p>We describe here a method to rapidly convert any desirable DNA fragment, as small as 100 bp, into long tandem DNA arrays up to 140 kb in size that are inserted into a microbe vector. This method includes rolling-circle phi29 amplification (RCA) of the sequence &l...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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BMC
2011-10-01
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| Series: | Biological Procedures Online |
| Online Access: | http://www.biologicalproceduresonline.com/content/13/1/8 |
| _version_ | 1811329334566191104 |
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| author | Kouprina Natalay Larionov Vladimir Lee Nicholas CO Noskov Vladimir N |
| author_facet | Kouprina Natalay Larionov Vladimir Lee Nicholas CO Noskov Vladimir N |
| author_sort | Kouprina Natalay |
| collection | DOAJ |
| description | <p>Abstract</p> <p>We describe here a method to rapidly convert any desirable DNA fragment, as small as 100 bp, into long tandem DNA arrays up to 140 kb in size that are inserted into a microbe vector. This method includes rolling-circle phi29 amplification (RCA) of the sequence <it>in vitro </it>and assembly of the RCA products <it>in vivo </it>by homologous recombination in the yeast <it>Saccharomyces cerevisiae</it>. The method was successfully used for a functional analysis of centromeric and pericentromeric repeats and construction of new vehicles for gene delivery to mammalian cells. The method may have general application in elucidating the role of tandem repeats in chromosome organization and dynamics. Each cycle of the protocol takes ~ two weeks to complete.</p> |
| first_indexed | 2024-04-13T15:42:04Z |
| format | Article |
| id | doaj.art-569d793ceb024d0da501d56fe28baa27 |
| institution | Directory Open Access Journal |
| issn | 1480-9222 |
| language | English |
| last_indexed | 2024-04-13T15:42:04Z |
| publishDate | 2011-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Biological Procedures Online |
| spelling | doaj.art-569d793ceb024d0da501d56fe28baa272022-12-22T02:41:07ZengBMCBiological Procedures Online1480-92222011-10-01131810.1186/1480-9222-13-8Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomesKouprina NatalayLarionov VladimirLee Nicholas CONoskov Vladimir N<p>Abstract</p> <p>We describe here a method to rapidly convert any desirable DNA fragment, as small as 100 bp, into long tandem DNA arrays up to 140 kb in size that are inserted into a microbe vector. This method includes rolling-circle phi29 amplification (RCA) of the sequence <it>in vitro </it>and assembly of the RCA products <it>in vivo </it>by homologous recombination in the yeast <it>Saccharomyces cerevisiae</it>. The method was successfully used for a functional analysis of centromeric and pericentromeric repeats and construction of new vehicles for gene delivery to mammalian cells. The method may have general application in elucidating the role of tandem repeats in chromosome organization and dynamics. Each cycle of the protocol takes ~ two weeks to complete.</p>http://www.biologicalproceduresonline.com/content/13/1/8 |
| spellingShingle | Kouprina Natalay Larionov Vladimir Lee Nicholas CO Noskov Vladimir N Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes Biological Procedures Online |
| title | Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes |
| title_full | Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes |
| title_fullStr | Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes |
| title_full_unstemmed | Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes |
| title_short | Rapid generation of long tandem DNA repeat arrays by homologous recombination in yeast to study their function in mammalian genomes |
| title_sort | rapid generation of long tandem dna repeat arrays by homologous recombination in yeast to study their function in mammalian genomes |
| url | http://www.biologicalproceduresonline.com/content/13/1/8 |
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