BMB 472 Experimental Biochemistry

Experimental Approach to Molecular Biochemistry

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BMB 472 Learning Modules

Growth of bacteria monitored by spectrophotometry.

Growth of bacteriophage, titering, and transduction.

Preparation of bacteriophage and plasmid DNA.

Electrophoretic Analysis of DNA preps.

Directional cloning of topAcysB into pTrc99A.

Transformation of DM700 and XL-1 Blue with pTrc99A/topAcysB

Electrophoretic Analysis of Restricted DNAs and Products of Ligation

PCR Amplification of DNA

Analysis of Topoisomerase I Assays

Ligation of topA DNA into M13mp19

Densitometric Analysis of SDS-PAGE Gels and Western Blots

Transfection of F' E. coli cells with the Ligation Mixture of topA DNA Inserted into M13mp19

Orientation Analysis of topA Subclones by a Complimentarity Test