PCR-based Chromosome Walking: The SFR 3 Case
نویسندگان
چکیده
منابع مشابه
Chromosome walking by cloning of distinct PCR fragments.
For functional characterization of genes, it is often interesting to identify additional upor downstream sequences in a fast and inexpensive way. Constructing a phage λ or cosmid library is time consuming, and subcloning of the corresponding fragments has to be performed to isolate DNA that can be sequenced (3). Alternative methods such as inverse PCR (2) or onesided PCR (4) have been developed...
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A chromosome walking method was improved in this work. The new method was named as nonspecific primer anchored PCR (NPA-PCR). Nested gene specific primers were designed based on the known region and long random primer using degeneracy oligonucleotides for nonspecific anchoring. Annealing temperatures were varied to control the priming. Target sequences were obtained by PCR with random primer an...
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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...
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Current genome walking methods are cumbersome to perform and can result in non-specific products. Here, we demonstrate the use of partially overlapping primer-based PCR (POP-PCR), a direct genome walking technique for the isolation of unknown flanking regions. This method exploits the partially overlapping characteristic at the 3' ends of a set of POP primers (walking primers), which guarantees...
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We developed a self-formed adaptor PCR (termed SEFA PCR) which can be used for chromosome walking. Most of the amplified flanking sequences were longer than 2.0 kb, and some were as long as 6.0 kb. SEFA PCR is simple and efficient and should have broad applications in the isolation of unknown sequences in complex genomes.
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ژورنال
عنوان ژورنال: Pakistan Journal of Biological Sciences
سال: 2005
ISSN: 1028-8880
DOI: 10.3923/pjbs.2006.164.169