August/7 August 2009

From 2009.igem.org

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Latest revision as of 07:34, 12 October 2009

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)

            [http://partsregistry.org/wiki/index.php?title=Part:BBa_F1610 F16010]  (2-24G)                        none
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_M0100 M0100]   (2-15F)                        none
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_I742158 I742158] (1-19B)                     >50
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0034 B0034]   (1-2M)                         >50
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_I0462 I0462]   (1-8O)                         ~10
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118002 K118002] (2-16F)                     ~10
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118003 K118003] (2-16H)                     ~10
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0015 B0015]  (1-23L)                         >50
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_I1466 I1466]  (1-23J)                         ~10
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0077 C0077]  (1-14F)                        ~10
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0076 C0076]  (1-14D)                        ~10
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_K081020 K081020] (2-12H)                     none
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_S03878 S03878]  (2-16M)                      ~10
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_C0179 C1079]   (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 compe[tent cells with the following parts to amplify them:

(plate number)-(location on plate) (antibiotic resistance)

            [http://partsregistry.org/wiki/index.php?title=Part:BBa_F1610 F16010]  (2-24G)                             A
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_K081020 K081020] (2-12H)                             K
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_K118013 K118013] (2-18F)                             A
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0071 R0071] (1-12C)                             A
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_I14017 I14017] (1-18L)                             K
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0062 R0062] (1-6O)                              A
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0079 R0079] (1-12A)                             A
            [http://partsregistry.org/wiki/index.php?title=Part:BBa_R0077 R0077] (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).

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