MBIO 3410 Lecture Notes - Lecture 2: Ethidium Bromide, Nucleic Acid Thermodynamics, Gc-Content
24 Dec 2015
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Cut up dna into small bits (40 bp), denature, and slowly cool. Highly repetitive sequences will reanneal more quickly than random or less repetitive sequences. Dna melting curve: tm: 50% denaturation. Repetitive dna should reassociate more quickly compared to unique: probability of two complementary strands meeting is higher. Cot curves: concentration x incubation time: measure concentration and genome complexity, viruses overall renature the fastest simplest genomes. Highly repetitive dna found around telomere and centromere. Middle repetitive dna rdna (ribosomal genes for production of ribosomes), certain families of mobile dna. Single copy non-repetitive; protein coding genes, unique (cid:1) (cid:1) (cid:1) regions sequences electrode. Separation of dna fragments based on molecular weight. Dna/rna carry a negative charge, so they move towards a positive. Agarose or polyacrylamide gels contain pores that allow small molecules to pass through. Visualize dna by staining gel with ethidium bromide.
