Selective Isolation of Mammalian Genes by <scp>TAR</scp> Cloning

Kouprina, Natalay; Larionov, Vladimir

doi:10.1002/0471142905.hg0517s49

Cited by 4 publications

(1 citation statement)

References 29 publications

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“…We found a strategy to clone large fragments of DNA by dividing them to short PCR fragment cloning, followed by restriction digestion and ligation cloning 9 . This strategy shows dominance in amplifying efficiency, it helps overcome PCR problem, and constructs very big clones without turning to complex reagents and technologies, like Red/ET system 10 or TAR system 11 , 12 . This strategy shows potential in handling long CDS clones, including introducing mutation, deleting and inserting domains.…”

Section: Introductionmentioning

confidence: 99%

Restriction-based Multiple-fragment Assembly Strategy to Avoid Random Mutation during Long cDNA Cloning

Wang¹,

Chen²,

Zhang³

et al. 2015

J. Cancer

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Long fragment cloning is a challenge for its difficulty in accurate amplifying and tendency to get unwanted mutation. Here we discuss Restriction-based Multiple-fragment Assembly Strategy's advantages and limitations. In this strategy, rather than PCR amplifying the entire coding sequence (CDS) at one time, we amplified and sequenced smaller fragments which are shorter than 1.5kb spanning the CDS. After that, the sequence-proved fragments were assembled by digestion-ligation cloning to the target vector. We test its universality in our script programmed in Python. Our data shows that, among the entire human and mouse CDS, at least 70% of long CDS cloning will benefit from this strategy.

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Section: Introductionmentioning

confidence: 99%

Restriction-based Multiple-fragment Assembly Strategy to Avoid Random Mutation during Long cDNA Cloning

Wang¹,

Chen²,

Zhang³

et al. 2015

J. Cancer

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Transformation-Associated Recombination (TAR) Cloning of Tumor-Inducing Xmrk2 Gene from Xiphophorus maculatus

Kirchner

Ivanova

Samson

et al. 2001

Marine Biotechnology

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We modified the TAR methodology of YAC clone construction for application to fish genomic DNA isolated from Xiphophorus maculatus. YAC libraries were developed using the XIR1 repeat sequence as the recombinational hook. Construction of these libraries demonstrates that Xiphophorus DNA sequences can function as hooks in the yeast recombination system and that X. maculatus genomic DNA contains sequences that provide origin of replication function in yeast. By screening a subset of Xiphophorus YAC clones, we isolated a clone harboring the Xmrk2 locus that is associated with spontaneous and induced melanomagenesis.Modifications of the TAR technique allowed the targeted cloning of specific genes from genomic regions ranging in size from cDNAs to several hundred kilobases. Specific genomic regions can be isolated in a directional manner from fixed map locations to saturate these areas with physical markers. We discuss the applications of these and other yeast recombinational processes to fish genetics.

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Characterization of the hamster genomic fragment cloned by TAR cloning technology with interspecific sequence information

et al. 2012

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Selective Isolation of Mammalian Genes by TAR Cloning

Cited by 4 publications

References 29 publications

Restriction-based Multiple-fragment Assembly Strategy to Avoid Random Mutation during Long cDNA Cloning

Restriction-based Multiple-fragment Assembly Strategy to Avoid Random Mutation during Long cDNA Cloning

Transformation-Associated Recombination (TAR) Cloning of Tumor-Inducing Xmrk2 Gene from Xiphophorus maculatus

Characterization of the hamster genomic fragment cloned by TAR cloning technology with interspecific sequence information

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