SSCP Analysis Questions and Answers Rated A+

SSCP Analysis Questions and Answers Rated A+ What Does SSCP analysis stand for? Single strand conformation polymorphism analysis What is the simplest and most used method of mutation detection? SSCP analysis Give a simple summary of SSCP analysis procedure PCR is used to amplify the region of interest and the resultant DNA is separated as single stranded molecules by electrophoresis gel. On what premises does this whole procedure rely on? 1. Single strand DNA folds differently if it differs by a base. 2. Mutations induce changes in tertiary structure 3. Changes in tertiary structure result in different mobilities What 3 methods are used to detect the mutations? Radioactive detection, staining, fluorescent PCR pairs. Is it possible to detect whether a mutation will be detected under given conditions? Although the tertiary structure of single stranded DNA changes under different physical conditions, it is not possible What is the mutation detection sensitivity of PCR-SSCP? 80% in a single run for fragments shorter than 300bp T or F: The absence of a new band does not prove that there is no mutation in the analysed molecule. True, sensitivity is 80% not 100% what is the optimum fragment length for the sensitivity of PCR-SSCP? <300 bp, as fragment length increases, sensitivity decreases. cDNA ? DNA synthesized by reverse transcriptase. Reverse transcriptase? RNA-dependent DNA polymerase in many retroviruses. -converts RNA template molecules into a DNA double helix. What is cDNA often used for? Cloning eukaryotic genes in prokaryotes. Advantages of SSCP? -works on a specified region -simple and fast Limitations to SSCP? -only tells you that there is a mutation. -not all mutations have detectable mobility