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August/7 August 2009
From 2009.igem.org
Today we carried out the following procedures (in rough chronological order):
1. Checked the cell cultures transformed and plated out yesterday (8/6) for colony formation and made an approximate count of the number of colonies.
(plate number)-(location on plate) (no. of colonies)
2-24G none
2-15F none
1-19B >50
1-2M >50
1-8O ~10
2-16F ~10
2-16H ~10
1-23L >50
1-23J ~10
1-14F ~10
1-14D ~10
2-12H none
2-16M ~10
2-8M ~10
2. Conducted a 'miniprep' to harvest and check concentration of the EpsE (molecular clutch) plasmids from 3 test tube cultures incubated since yesterday (8/6). Result of Nanodrop concentration check:
(test tube no.) [260/280] [260/230] (concentration)
1 2.14 1.65 37.6 ng/ul
2 2.07 1.95 62.9 ng/ul
3 1.95 1.33 60.6 ng/ul
3. Prepared kanamycin antibiotic solution from kanamycin sulfate and MilliQ water. 100 mg kanamycin sulfate + 10 ml MilliQ water -> 10 ml of 10 mg/ml kanamycin solution.
4. Transformed competent cells with the following parts to amplify them:
(plate number)-(location on plate) (antibiotic resistance)
2-24G A
2-12H K
2-18F A
1-12C A
1-18L K
1-6O A
1-12A A
1-10K A
Note: 2-24G and 2-12H were transformed yesterday but produced no colonies. 2-18F was marked for transformation yesterday but 2-15F was mistakenly transformed instead.
5. Treated a sample of EpsE-containing plasmids with EcoRI and SpeI restriction endonucleases to isolate and check length of amplified EpsE part. The plasmid-buffer-endonuclease mixture was incubated and left to react overnight (Incubation at 37 degrees Celsius began at 16:00).
