Techniques to manipulate dna Phenol/chloroform extraction Ethanol precipitation Concentration determination Gel Electrophoresis dna hybridization Se outhern Blotting Melting tempe eratures PCR
Techniques to manipulate DNA Phenol/chloroform extraction Ethanol precipitation Concentration determination Gel Electrophoresis DNA hybridization Southern Blotting Melting temperatures PCR
Example: Is the brca3 gene involved in Suppressing breast cancer? BRCAl BRCA3 BRCA2 EcoRl bRCA ECoRl NIH
EcoRI BRCA3 EcoRI NIH Example: Is the BRCA3 gene involved in Suppressing breast cancer? BRCA1 BRCA2 BRCA3
Southern blotting ETOH ppt neno ProtK SDS Chloro tissue 9 Cleave with restriction enzymes Nitrocellulose Autoradiography Paper Towels Hybridize with labeled DNA Filter paper Nitrocellulose RNA probe: perform esIS autoradiography Capillary action transfers DNA from gel to nitrocellulose
Paper Towels Southern Blotting 1 tissue Phenol ProtK SDS Chloro ETOH ppt Spin
The"Hyperchromic Shift 0.4 0.3 0.2 220 240 280nm
The “Hyperchromic Shift” 2
Southern blotting ETOH ppt neno ProtK SDS Chloro tissue 9 Cleave with restriction enzymes Nitrocellulose Autoradiography Paper Towels Hybridize with labeled DNA Filter paper Nitrocellulose RNA probe: perform esIS autoradiography Capillary action transfers DNA from gel to nitrocellulose
Paper Towels Southern Blotting 1 tissue Phenol ProtK SDS Chloro ETOH ppt Spin