Computation-assisted SiteFinding- PCR for isolating flanking sequence tags in rice
نویسندگان
چکیده
منابع مشابه
Cloning flanking sequence by single-primer PCR in transgenic plants.
The insertion position of exogenous genes in plant genomes is usually identified by adapter ligation-mediated polymerase chain reaction (PCR), thermal asymmetric interlaced PCR, and restriction site extension PCR in transgenic plant research. However, these methods have various limitations, such as the complexity of designing primers and time-consuming and multiple-step procedures. The goal of ...
متن کاملSiteFinding-PCR: a simple and efficient PCR method for chromosome walking
In this paper, we present a novel PCR method, termed SiteFinding-PCR, for gene or chromosome walking. The PCR was primed by a SiteFinder at a low temperature, and then the target molecules were amplified exponentially with gene-specific and SiteFinder primers, and screened out by another gene-specific primer and a vector primer. However, non-target molecules could not be amplified exponentially...
متن کاملGeneration of a flanking sequence-tag database for activation-tagging lines in japonica rice.
We have generated 47,932 T-DNA tag lines in japonica rice using activation-tagging vectors that contain tetramerized 35S enhancer sequences. To facilitate use of those lines, we isolated the genomic sequences flanking the inserted T-DNA via inverse polymerase chain reaction. For most of the lines, we performed four sets of amplifications using two different restriction enzymes toward both direc...
متن کاملA PCR-based method for isolation of genomic DNA flanking a known DNA sequence.
We describe a simple PCR-based method for the isolation of genomic DNA that lies adjacent to a known DNA sequence. The method is based on the directional cloning of digested genomic DNA into the multiple cloning site of a pUC-based plasmid to generate a limited genomic library. The library is plated onto a number of selective LA plates which are incubated overnight, and recombinant plasmid DNA ...
متن کاملExpressed Sequence Tags
Now a days, genome analysis has employed a rapid analysis tool known as expressed sequence tags. In 1983, SD Putney for the first time demonstrated the use of cDNA in identification of genome. The term expressed sequence tags (ESTs) was coined by Anthony Kerlavage at the Institute for Genomic Research. In 1991, Mark Adams used EST in relation to gene discovery and Human Genome project. cDNA or ...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 2011
ISSN: 0736-6205,1940-9818
DOI: 10.2144/000113787