Biochemistry 2288A Lecture Notes - Lecture 14: Apoptosis, Paclitaxel, Cell Adhesion
5 Oct 2017
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Nacl) to the solution do to the tm of a dna. Extrinsic factors that affect the tm: salt concentration, higher salt concentration= higher tm, positively charged ions (ex. Na+) shield the repulsive interactions of the negatively charged phosphate backbone. Lowers tm: solvents that disrupt h bonding interfere with base pairing and lower the tm. Melting is reversible: process called renaturation, hybridization or annealing, ***hybridization requires complementary strands. Hybridization will detect complementary nucleic acid sequences: hybridization are often done after transferring the nucleic acid to a membrane, southern blot: dna to dna on the membrane, northern blot: dna to rna on the membrane. Nucleic acid separation is based on size using gel electrophoresis. Positive, since dna at neutral ph is negatively charged. Gel electrophoresis: dna is sieved through a matrix of agarose being pulled by an electric field, agarose after melting, it hardens into a form like jello. When stained with ethidium bromide, dna fluoresces red under uv.
