Screening of differentially amplified cDNA products from RNA arbitrarily primed PCR fingerprints using single strand conformation polymorphism (SSCP) gels

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Screening of differentially amplified cDNA products from RNA arbitrarily primed PCR fingerprints using single strand conformation polymorphism (SSCP) gels.

Arbitrarily primed PCR fingerprinting of RNA and differential display resolved on an acrylamide gel has been extensively used to detect differentially expressed RNAs. However, after a differentially amplified product is detected the next steps are labor-intensive: a small portion of the fingerprinting gel that contains the differentially amplified product is cut out, reamplified and the correct...

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Arbitrarily primed PCR fingerprints resolved on SSCP gels.

Arbitrarily primed PCR methods (1), including RAPD (2), generate a reproducible fingerprint of DNA products from complex nucleic acids. Differences detected between fingerprints derived from different genomes have been used extensively as polymorphisms in genetic mapping (e.g., 3). Various strategies can be used to maximize the number of polymorphisms that are scored in each fingerprinting expe...

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non-radioactive single-strand conformation polymorphism (sscp) analysis of relatively long pcr products

the sscp technique is based on the appearance of new “refolding” conformations during electro-phoresis due to mutation. in order to develop a simple, non-radioactive sscp analysis method so that it can reliably detect single nucleotide changes in pcr products up to 500 bp in length, extensive optimisation trials were performed . the best separation of sscp bands of pcr products up to 500 bp in ...

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Screening for mutations by RNA single-strand conformation polymorphism (rSSCP): comparison with DNA-SSCP.

Single-strand conformation polymorphism (SSCP) is a simple method for detecting the presence of mutations in a segment of DNA, but the fraction of all mutations detected is unclear. We have evaluated SSCP for the detection of single-base mutations in the factor IX gene. Multiple conditions were examined including electrophoresis temperature, electrophoresis buffer concentration, acrylamide to b...

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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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ژورنال

عنوان ژورنال: Nucleic Acids Research

سال: 1996

ISSN: 1362-4962

DOI: 10.1093/nar/24.8.1504