Multiple Fluorescence-Based PCR-SSCP Analysis Using Internal Fluorescent Labeling of PCR Products
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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A fluorescence-based method for polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis, F-SSCP, was developed in which the target sequence is amplified by the PCR using fluorescent primers. The amplified products are then heat-denatured and applied to a water-jacket controlled gel in an automated DNA sequencer. The separated strands are detected as laser-excited f...
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amide gel electrophoresis (PAGE) on a 180× 160× 1.5-mm 10% polyacrylamide gel. The separated DNA strands were subsequently visualized by staining with either EtdBr or silver (2). Separation of the DNA strands before electrophoresis using streptavidin paramagnetic beads consistently yielded single-stranded products upon SSCP analysis (Figure 1). Treatment of DNA strands with S1 nuclease before S...
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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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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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ژورنال
عنوان ژورنال: BioTechniques
سال: 1996
ISSN: 0736-6205,1940-9818
DOI: 10.2144/96213rr05