Universal fluorescent labeling of PCR products for DHPLC analysis: reducing cost and increasing sample throughput
نویسندگان
چکیده
منابع مشابه
Multiple fluorescence-based PCR-SSCP analysis using internal fluorescent labeling of PCR products.
The method to internally label PCR products with multiple colored fluorescent dyes was developed and applied to multiple fluorescence-based PCR single-stranded conformational polymorphism (MF-PCR-SSCP) analysis. PCR-amplified fluorescent DNA fragments, which were internally labeled by adding fluorescent dUTPs ([F]dUTPs) to the PCR mixture, were heat-denatured and applied to a nondenaturing poly...
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The use of fluorescently labeled nucleotides (F-dUTP) in the PCR for genetic typing of microsatellite polymorphisms was investigated. Microdissected tumor cells from cervical squamous cancer biopsies were compared with cells from surrounding normal tissue for loss of heterozygosity (LOH) at the mismatch repair gene hMLH1. Removal of unincorporated F-dUTP before analysis is necessary to reduce b...
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A method for fluorescent postlabeling of PCR products has been developed. The method uses Klenow fragment of DNA polymerase I that exchanges the 3'-terminal residue of PCR-amplified DNA fragment for fluorescent nucleotides. All reactions, including PCR, are performed in one tube simply by successive addition of reagents. The products can be applied directly to fluorescence-based automated DNA s...
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1.Meldrum, D. 2000. Automation for genomics, part two: sequencers, microarrays, and future trends. Genome Res. 10:12881303. 2.Orlando, C., P. Pinzani, and M. Pazzagli. 1998. Developments in quantitative PCR. Clin. Chem. Lab. Med. 36:255-269. 3.Rudi, K., S.L. Flateland, J.F. Hanssen, G. Bengtsson, and H. Nissen. 2002. Development and evaluation of a 16S rDNA array approach for describing complex...
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A method for fluorescent labeling of PCR products has been developed. This method consists in a two-step procedure in which a first exponential classical PCR is followed by a "linear amplification". This second step relies on incorporation of fluorescent dNTP (dUTP or dCTP) in order to label the product on only one strand. The products can be applied without prior purification directly to a gel...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 2005
ISSN: 0736-6205,1940-9818
DOI: 10.2144/05391bm02