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From 2012.igem.org

I: Inoculate ECNR2 Cultures from Natalie Ma's Plate(from 7 Jun 2012)

• Inoculate one isolated colony into ~2 mL of LB media in sterile culture tube (x2)
• Place tubes in 37 ^oC and check on cultures after 4 hours to see if they have grown at all

• Results: No turbidity even after 4.5 - 5 hours in shaker. Suspected problem is that the toxic Gam protein of the recombination set is starting to be expressed at 37 ^oC. Cells for recombination protocol are normally grown at 30 ^oC, but can tolerate as high as 34 ^oC according to Professor Isaacs. Since the other cells should exhibit similar growth at 34 ^oC as at 37 ^oC, I will lower the shaking incubator temperature to 34 ^oC Friday morning.

• Incubate cultures overnight with shaking at 37 ^oC

• The cap came off one of the cultures, so it was discarded immediately to avoid contamination

II. Grown ECNR1/ECNR2 streaked plate (from Prof. Isaacs) overnight

• Place in the static incubator (~34 - 35 ^oC)