bac gap repair
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1Departments of Pediatrics and Genetics Cell Biology and
the precise isolation of large-size DNA fragments Unlike PCR amplification DNA gap repair (DGR) is a virtually error-free process However the maximal size of bacterial artificial chromosome (BAC) insert isolated so far by recombination-mediated genetic engineering (recombineering) is |
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P[acman]: A BAC Transgenic Platform for Targeted Insertion of
artificial chromosome (BAC) recombineering and φC31-mediated transgenesis The BAC is maintained at low-copy number facilitating plasmid maintenance and recombineering but induced to high-copy number for plasmid isolation Recombineering allows gap repair and mutagenesis in bacteria Gap repair efficiently retrieves |
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Simple and highly efficient BAC recombineering using galK
fragment from a BAC by gap repair either for use as a transgene directly or for subsequent manipulation to make a gene-targeting construct The introduction of selectable mark-ers into a BAC is also very easy using recombineering How-ever a major limitation to the usefulness of BACs is the ease |
How is P[Acman] used to retrieve Frag- ments by Gap Repair?
P[acman] was used to retrieve frag- ments by gap repair. For each gap repair, we designed four primer sets (fig. S1). Two ho- mology arms, located at either end of the DNA fragment, were cloned into P[acman] (fig. 1B).
Is plasmid-based gap-repair a reliable method for recombineering in E coli?
Using recombineering in E. coli, we demonstrate that plasmid-based gap-repair is a highly efficient and reliable method to introduce mutations into large plasmids circumventing the time-consuming and tedious introduction of selection cassettes and counter selection for desired mutations during manipulation of BACs or other large vectors.
Is gap-repair recombineering useful in PhiC31 integrase-mediated transgenesis?
We then developed gap-repair recombineering to very efficiently manipulate these large reporter plasmids in Escherichia coliusing restriction enzymes or sgRNA/Cas9 DNA scission to capitalize on the many benefits of plasmids in phiC31 integrase-mediated transgenesis.
How does gap repair recombineering work?
The high efficiency of DNA manipulation by gap-repair recombineering requires introducing two double-strand cuts into the DNA. Generally, restriction enzymes can be identified that cut only once within a 20 kb plasmid, but such sequence limitations can now be overcome by the use of the sgRNA/Cas9 endonuclease.
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Isolation of Full Size BAC Inserts by DNA Gap Repair in E. coli
14-Feb-2019 Unlike PCR amplification DNA gap repair. (DGR) is a virtually error-free process. However |
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Recombineering & Gap Repair Protocol
retrieved using gap repair and turned into a gene targeting vector. The BAC modifications are done using a modified bacterial strain SW102. |
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P[acman]: A BAC Transgenic Platform for Targeted Insertion of
30-Nov-2006 Gap repair efficiently retrieves DNA fragments up to 133 kilobases long from P1 or BAC clones. fC31- mediated transgenesis integrates these ... |
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P[acman]: A BAC Transgenic Platform for Targeted Insertion of
30-Nov-2006 Gap repair efficiently retrieves DNA fragments up to 133 kilobases long from P1 or BAC clones. fC31- mediated transgenesis integrates these ... |
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P[acman]: A BAC Transgenic Platform for Targeted Insertion of
Recombineering allows gap repair and mutagenesis in bacteria. Gap repair efficiently retrieves DNA fragments up to 133 kilobases long from P1 or BAC clones. |
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DOI: 10.1126/science.1134426 1747 (2006); 314 Science et al
04-Jan-2007 Gap repair efficiently retrieves DNA fragments up to 133 kilobases long from P1 or BAC clones. fC31- mediated transgenesis integrates these ... |
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BACâ•recombineering for studying plant gene regulation
14-Jan-2011 which are suitable for gap-repair cloning of genes from BACs using recombineering ... Modification of large DNA inserts of BAC and P1 (PAC). |
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University of Birmingham Plasmid-based gap-repair recombineered
12-Dec-2018 coli (recombineering) provides a versatile alternative to manipulate DNA par- ticularly in large Bacterial Artificial Chromosomes (BACs) and ... |
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Plasmid-based gap-repair recombineered transgenes reveal a
coli (recombineering) provides a versatile alternative to manipulate DNA par- ticularly in large Bacterial Artificial Chromosomes (BACs) and viruses (17–21) |
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Plasmid-based gap-repair recombineered transgenes reveal a
12-Dec-2018 coli (recombineering) provides a versatile alternative to manipulate DNA par- ticularly in large Bacterial Artificial Chromosomes (BACs) and ... |
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Isolation of Full Size BAC Inserts by DNA Gap Repair - bioRxiv
Cité 1 fois — When a gap is present in a cloning vector, DGR will allow a new DNA sequence to be |
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Recombineering & Gap Repair Protocol
retrieved using gap repair and turned into a gene targeting vector The BAC The protocol proposed uses electroporation to transfer the BAC DNA into SW102 Alternatively |
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P[acman]: A BAC Transgenic Platform for Targeted Insertion of
Cité 726 fois — Gap repair efficiently retrieves DNA fragments up to 133 kilobases long from P1 or BAC clones fC31- |
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P[acman]: A BAC Transgenic Platform for Targeted - Science
2006 · Cité 726 fois — Gap repair efficiently retrieves DNA fragments up to 133 kb from P1 or BAC clones |
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P[acman]: A BAC Transgenic Platform for Targeted - Science
content › sci |
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Review Article Bacterial Artificial Chromosome Mutagenesis
Cité 36 fois — (d) Gap repair cloning Figure 1: General applications of recombineering in BAC modifications |
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Copeland-2001 Nat rev Genetpdf - RECOMBINEERING
2001 · Cité 908 fois — BAC DNA Regions of homology Target gene Figure 4 Retrieving cloned DNA by gap repair |
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Lambda Red Mediated Gap Repair Utilizes a Novel - AWS
2015 · Cité 5 fois — cassette into the BAC followed by gap repair (B) Consecutive route 2; first gap repair |
