斯坦福大学分子生物学讲义--DNAII.pptVIP

Techniques to manipulate DNA PolyAcrylamide Gels What parameters affect theTm?(Tm is defined as the temp at which half the moleculesare single-stranded) Kinetics of Hybridization:Do I have time for a soda or a week-end in New York City? Results of Southern Blot Results of PCR #1 Results of PCR#5 PCR mutagenesis EcoRI HindIII Isolate products, mix Amplify digest, subclone sequence 20 PvuII * Phenol/chloroform extraction Ethanol precipitation Concentration determination Gel Electrophoresis DNA hybridization Southern Blotting Melting temperatures PCR BRCA3 EcoRI EcoRI NIH Example: Is th