BIOL309 Study Guide - Final Guide: Alkaline Phosphatase, Caesium Chloride, Molecular Cloning

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Denaturation: separation of two strands of nucleic acid that only h bonds holding strands together are disrupted and other bonds not affected. Treatments that lead to denaturation: low salt, high temperature thermal energy break h bonds but not covalent linkages of backbone, high ph, formaldehyde urea base pair with bases or h-bond groups. Hypochromic shift: shift from denatured state back to double strand. Nucleic acid synthesis to 3" direction using ribonucleotide triphosphates (ntps) Incoming dntp added to 3" oh group of last nucleotide of daughter strand. All strands extended in 5" to 3" direction. Dna polymerases require primer - source of 3" oh group. Rna polymerases do not need primer but synthesize ribose nucleotide strands in 5" Dna to be cloned fragmented with restriction enzymes. Two dnas joined using t4 dna ligase and introduced into host cells. Reduce hydrophobic effect of bases -> contribute to dna stability. In vitro dna synthesis reaction: homogenize tissue and lyse cells.

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