Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/23076
Title: Target quantitative detection and discrimination study by quenching probes, molecular beacons and quenching beacons
Authors: RONG YAN
Keywords: hybridization, discrimination, melting temperature, oligonucleotide probe, perfect match, single base mismatch
Issue Date: 18-May-2007
Source: RONG YAN (2007-05-18). Target quantitative detection and discrimination study by quenching probes, molecular beacons and quenching beacons. ScholarBank@NUS Repository.
Abstract: Quenching probes (QP), molecular beacons (MB), and quenching beacons (QB) were investigated for their ability to quantitatively detect specific DNA target and discriminate single nucleotide mismatch (MM). We designed QB based on the structure of MB. Two sets of probes were also designed, each of which included a perfect match (PM) target and 54 targets with single MM of all types at all positions. Hybridization was carried out in homogeneous solution. Fluorescence quenching efficiencies and signal-to-background (S:B) ratios of the probes were discussed. The differences in melting temperatures between PM duplex and MM duplexes (delta Tm) were exploited. Results suggested that BODIPY FL- or TAMRA-labeled QPs were good candidates for quantitative detection of nucleic acid. QBs showed reduced S:B ratio and specificity compared with MBs. Many MMs could be successfully discriminated with delta Tm>2A?C. Both MM position and MM type were shown to significantly affect discrimination of single base MM.
URI: http://scholarbank.nus.edu.sg/handle/10635/23076
Appears in Collections:Master's Theses (Open)

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