BCHM 4116 1st Edition Lecture 8 Outline of Last Lecture I. Gibson assembly II. Recombination III. DNA lib.IV. Genome lib. Outline of Current Lecture I. HybridizationCurrent LectureI. Hybridization: Southern blot, northern blot & lib screeninga. Hybridizationi. Formation of a ds duplex nucleic acid moleculeii. When one strand is labeled and used as probeiii. This technique allows the detection of nucleic acid molecules that are complementary to the known probe b. Blottingi. Transfer of DNA or RNA from gel to filter c. Southern bloti. DNA separated via gele electrophoresisii. Resulting fragments transferred (blotted) to solid support (membrane or filter) iii. A labeled probe is used to locate the position of complementary sequence by hybridization iv. Factors that affect southern blotting1. Stringency = how close a match b/w strands is required for duplex formation maintaineda. High = PICKYb. Less = less picky 2. Stability of probe measured by its Tm a. High Tm = more stable i. Therefore low stringency b. Low tm = less stablei. Therefore needs to be high stringency so that probe detects right one d. Northern blotThese notes represent a detailed interpretation of the professor’s lecture. GradeBuddy is best used as a supplement to your own notes, not as a substitute.i. Same as southern but for
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