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From 2010.igem.org

Contents 1 Nina 1.1 Preparation of competent DH5α Escherichia coli cells 2 Johan 2.1 Continuation of making competent cells

Nina

Preparation of competent DH5α Escherichia coli cells

Continued from 21/6.

Prepared competent DH5α cells according to the protocol of Morten Nørholm at the Department of Biochemistry & Biophysics Stockholm University.

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Absorbance:

• 1 ml sample from one bottle:

Time				OD
30min				0,029
1h				0,053
1h				0,188
30min				0,316

• 1 ml sample from two bottles:

Time				OD
30min				0,519/0,503
7min				0,578/0,566
5min				0,598/0,600

Johan

Continuation of making competent cells

Step 2-12 in "800 ml of competent cells"

1 L instead of 800 ml -> 25 st 50 ml Falcon Tubes with 40 ml, 1 st with 15-20 ml

10) 16/23 = 0,69 ml

3) Abs:

Only one bottle

• 0,029 9:25
• 0,053 ~1h5min
• 0,188 ~1h5min
• 0,316 ~30min

Both bottles

• 0,519 ~30min
• 0,503
• 0,578 ~7min
• 0,566
• 0,598 ~5min = 13:05
• 0,600

12) ~160 epps, 100 µl in each.

They were then all put in the freezer, in two cardboard boxes (no metal blocks used)