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 Hybridization Current Lecture I Hybridization Southern blot northern blot lib screening a Hybridization i Formation of a ds duplex nucleic acid molecule ii When one strand is labeled and used as probe iii This technique allows the detection of nucleic acid molecules that are complementary to the known probe b Blotting i Transfer of DNA or RNA from gel to filter c Southern blot i DNA separated via gele electrophoresis ii 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 blotting 1 Stringency how close a match b w strands is required for duplex formation maintained a High PICKY b Less less picky 2 Stability of probe measured by its Tm a High Tm more stable i Therefore low stringency b Low tm less stable i Therefore needs to be high stringency so that probe detects right one d Northern blot These 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 RNA
View Full Document
Unlocking...