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Ligation-mediated PCR: uses small DNA linkers ligated to the DNA of interest and multiple primers annealing to the DNA linkers; it has been used for DNA sequencing, genome walking, and DNA footprinting Legacy Family Tree Deluxe 6. Legacy Family Tree Deluxe 6 - It may be performed manually by heating the reaction components to the melting temperature (e. DNA helicase, an enzyme that unwinds DNA, is used in place of thermal denaturation. Legacy Family Tree Deluxe 6 - It involves a series of DNA digestions and self ligation, resulting in known sequences at either end of the unknown sequence. Hot-start/cold-finish PCR is achieved with new hybrid polymerases that are inactive at ambient temperature and are instantly activated at elongation temperature. Legacy Family Tree Deluxe 6 - Intersequence-specific PCR (ISSR): a PCR method for DNA fingerprinting that amplifies regions between simple sequence repeats to produce a unique fingerprint of amplified fragment lengths. Legacy Family Tree Deluxe 6 - Methylation-specific PCR (MSP): developed by Stephen Baylin and Jim Herman at the Johns Hopkins School of Medicine, and is used to detect methylation of CpG islands in genomic DNA. Legacy Family Tree Deluxe 6 - Because of the slow (arithmetic) amplification later in the reaction after the limiting primer has been used up, extra cycles of PCR are required. Legacy Family Tree Deluxe 6 - Helicase-dependent amplification: similar to traditional PCR, but uses a constant temperature rather than cycling through denaturation and annealing/extension cycles. Legacy Family Tree Deluxe 6 - Inverse PCR: is commonly used to identify the flanking sequences around genomic inserts. Legacy Family Tree Deluxe 6 - Hot-start PCR: a technique that reduces non-specific amplification during the initial set up stages of the PCR. Legacy Family Tree Deluxe 6 - , 95°C) before adding the polymerase.