Beta-Gal Induction Lab Report

Beta-Gal Induction Lab Report - INDUCTION OF -GALACTOSIDASE...

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INDUCTION OF β-GALACTOSIDASE Objective: In this lab we measured the induction rate of lac operon by, by measuring the β- Galactosidase produced. Methods: We were provided with E. coli that was grown in glucose medium at 37°C to a stationary phase and then diluted back to early log phase with glycerol 90 minutes prior to use. Solution was kept in shaking water bath to ensure that the solution stays well mixed (91). We added 0.5 mL of 2.0 mM IPTG (isopropylthiogalactopyranoside) to 5.0 mL of E. coli . We used IPTG to induce lactose operon instead of lactose. IPTG is a better inducer than lactose because it enters a cell at much faster rate. Lactose requires production of permease to open up the cell membrane to let in more lactose, IPTG does not permease, therefore there is no long induction lag when IPTG is used instead of lactose. Even if we had time to use lactose as the inducer, it would be undesirable to do so because lactose is substrate of β-Galactosidase. Afterwards we used 0.1 mL this induced culture and added it to 0.9 mL or reducing
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This note was uploaded on 04/30/2009 for the course BIOLOGY 037875 taught by Professor Athwal during the Spring '09 term at Temple.

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Beta-Gal Induction Lab Report - INDUCTION OF -GALACTOSIDASE...

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