August/7 August 2009

From 2009.igem.org

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===design genetic sercuit===
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Today we carried out the following procedures (in rough chronological order):
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*Todays work
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**build basic genetic circuit and make a parts list
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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.
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(plate number)-(location on plate)  (no. of colonies)
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-----------------------------------------------------------
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            2-24G                        none
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            2-15F                        none
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            1-19B                        >50
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            1-2M                        >50
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            1-8O                        ~10
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            2-16F                        ~10
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            2-16H                        ~10
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            1-23L                        >50
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            1-23J                        ~10
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            1-14F                        ~10
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            1-14D                        ~10
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            2-12H                        none
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            2-16M                        ~10
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            2-8M                        ~10
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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).
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Result of Nanodrop concentration check:
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(test tube no.) [260/280] [260/230] (concentration)
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---------------------------------------------------
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      1          2.14      1.65    37.6 ng/ul
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      2          2.07      1.95    62.9 ng/ul
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      3          1.95      1.33    60.6 ng/ul
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3. Prepared kanamycin antibiotic solution from kanamycin sulfate and MilliQ water.
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100 mg kanamycin sulfate + 10 ml MilliQ water -> 10 ml of 10 mg/ml kanamycin solution.
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4. Transformed competent cells with the following parts to amplify them:
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(plate number)-(location on plate)  (antibiotic resistance)
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-----------------------------------------------------------
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            2-24G                            A
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            2-12H                            K
 +
            2-18F                            A
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            1-12C                            A
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            1-18L                            K
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            1-6O                              A
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            1-12A                            A
 +
            1-10K                            A
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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.
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 +
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).

Revision as of 16:35, 7 August 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) 

            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).

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