Category:Protocols: Difference between revisions

Revision as of 23:50, 20 December 2016

This category has the following 22 subcategories, out of 22 total.

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-Creating a E.Coli Stock Culture
-. Make LB media as per the protocol titled Media Preparation
-. Transfer 25mL of LB media in to a sterile (autoclave) 250 mL  erlenmeyer flask using a sterile Falcon Transfer Pipet
-. Inoculate the flask with enough stock culture(Glycerol stock obtained from a lab mate/purchased) to create a final OD600 of 0.025
-. Make note to have 1:10 ratio of liquid to air in your inoculated flask for aerobic growth of E.coli
-. Incubate the flask at 37C and a shaker speed of 100-150 rpm
-. Over every 30 min take an OD600 reading to track the growth of the E.coli cells in your flask
-. When the cells hit Stationary phase (OD600 greater than or equal to 10 in 1ml of culture) the culture is ready to make stock tubes for storage
-. At this point, take 1ml of culture and add 0.5ml of glycerol in a 2ml sterile tube, create 10-15 of these 2ml glycerol stock tubes
-. Store the tubes with appropriate labeling in the -80C freezer for until you need to use them again.