User:Michael F. Nagle/Notebook/Chem 571/2012/09/18: Difference between revisions
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#Starter Cultures with 25mL water had .8793g, .8800g, .8748g, and .8757g broth in them. | #Starter Cultures with 25mL water had .8793g, .8800g, .8748g, and .8757g broth in them. | ||
#Expression cultures with 1L water had 25.0289g, 25.0069g, 25.002g, and 25.0036g broth in them. | #Expression cultures with 1L water had 25.0289g, 25.0069g, 25.002g, and 25.0036g broth in them. | ||
#Amount of Kanamycin for cultures was calculated. The molarity was converted to µg/mL and the equation M<sub>1</sub>V<sub>1</sub> = M<sub>2</sub>V<sub>2</sub> was used to calculate the volume of stock that should go in broth. | #Amount of Kanamycin for cultures was calculated. Kanamycin is an antibiotic and is being used to prevent microbial growth besides that of E. Coli. | ||
##The molarity was converted to µg/mL and the equation M<sub>1</sub>V<sub>1</sub> = M<sub>2</sub>V<sub>2</sub> was used to calculate the volume of stock that should go in broth. | |||
##50mg/mL Kanamycin * 1µg/.001mg = 50,000µg/mL Kanamycin | ##50mg/mL Kanamycin * 1µg/.001mg = 50,000µg/mL Kanamycin | ||
##50µg/mL * 35mL = 50000µg/mL | ##50µg/mL * 35mL = 50000µg/mL * volume needed | ||
##.035mL Kanamycin | ##.035mL Kanamycin solution needed | ||
##The antibiotic was allowed to warm to room temperature and then inserted to the broth. The remaining Kanamycin went back in the freezer. | ##The antibiotic was allowed to warm to room temperature and then inserted to the broth. The remaining Kanamycin went back in the freezer. | ||
#The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time. | #The foil caps on the broth flasks was sterilized with a bunsen burner after each time they were removed and placed back on the flasks. Metal forceps, which were also sterilized with flame between uses, were used to pick up a splint, which was used to pick up one colony of E. Coli and transfer it to the broth. A different split was used each time. | ||
# | #E. Coli transformed with a plasmid coding for Adenosine Deaminase (ADA)was inserted into the broth. | ||
#The flasks were | #The flasks were left overnight in an orbital incubator shaker at 37°C and 237 rpm | ||
==Discussion== | |||
*ADA is being synthesised for upcoming experiments on it's potential ability to make gold into nanoparticles. | |||
Revision as of 17:44, 7 December 2012
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Objectives
ProcedureStarter Culture and Expression Culture
Discussion
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