BIOC 3400 Study Guide - Midterm Guide: Autoradiograph, Chemiluminescence, Klenow Fragment

• School: Dalhousie University
• Department: Biochem & Molecular Biology
• Course: BIOC 3400
• Professor: Liu Paul

Document Summary

Study online at quizlet. com/_1nwuod: denaturation / melting: the process of separation of double, what are the factors that affect tm? (factors affect hybrid stability): - base composition ((g+c)/(t+a) ratio): tm increases with increased g+c content. Degree of sequence similarity between hybrids: every 1% mismatching of bases in a dna duplex reduces the. Temperature: the reaction temperature affects the rate of nucleic acid hybridization. : - a method used in molecular biology for detection of a specific dna sequence in dna samples. *molecules labeled with fluorophores (dyes) with defferent emission wavelengths can be detected with film or fluorescence detector. Primers dna labeling system: a method to radioactively label dna; (a dna fragment is denatured by heating in a boiling water bath. Then, random sequence oligonucleotides are annealed to both strands. Klenow fragment polymerase is then used to extend the oligonucleotides, using three cold nucleotides and one radioactively labeled nucleotide provided in the reaction mixture, to produce a uniformly labeled double-stranded probe.