BISC 2539 Lecture Notes - Lecture 10: Recombinant Dna, Plasmid, Chromosome
24 Dec 2019
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Dnaamplification production ofmultiplecopiesof a dnasequence in vivo livebacterialcells or vitroctesttube vitro starts w asampleofdnamolecule donordna_ Vectorsfragmentof donordna insertedinto a speciallydesigned plasmidor bacterialvirusthat will carry and amplifygene ofinterest. Polymerasechainreactioncpcrd i a specificgene or dnaregion of interest is isolated and amplifiedbydnapolymerase and bind primers to it. 2 adnaligation design constructdnamoleculesthat are not found in nature. 4bnudei g cloningvectors1pcr portion of dnarepeatedlycopied togeneratecopies restrictionnuclease. Hybridization find specificsequence of dnarna w great accuracy abilitytoselectivelybind to complementarynucleic acid sequences. 5 bdnaseguencing rapiddetermination of the sequenceof nucleotides of anydnaceven entiregenome. Isolatesinglegene pcramplifyselectedregions of dna vitro c codingsequence ofgene w o introns cdnacdnacopies of mrnaproducts can besynthesized inserted into vectors. Recut atspecific restrictions restrictionfracgments determinedbylocation of restriction segment dnapalindrome samenucleotide antiparallel types. Dnapolymerseadd bases to the 3 end ofprimer copysequence. 2 cooling at 50 650c single strand anheal to complementarystrand. 3 heat tolerant dnaboly replicates to single stranded extending from primer.
