E-Notebook
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| National Centre for Biological Sciences, Bangalore |
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| Bangalore | The Company | The Mission | e-Notebook |
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| The Bangalore iGEM Journal, 07
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Contents |
e-notebook
| May 07 | June 07 | July 07 | ||||||||||||||||||||||||||||||||
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| 1 | 2 | 3 | 4 | 5 | 1 | 2 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | |||||||||||||||||||||
| 6 | 7 | 8 | 9 | 10 | 11 | 12 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | ||||||||||||||
| 13 | 14 | 15 | 16 | 17 | 18 | 19 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | 15 | 16 | 17 | 18 | 19 | 20 | 21 | ||||||||||||||
| 20 | 21 | 22 | 23 | 24 | 25 | 26 | 17 | 18 | 19 | 20 | 21 | 22 | 23 | 22 | 23 | 24 | 25 | 26 | 27 | 28 | ||||||||||||||
| 27 | 28 | 29 | 30 | 31 | 24 | 25 | 26 | 27 | 28 | 29 | 30 | 29 | 30 | |||||||||||||||||||||
Current Status
Tasks Accomplished
Experiments so far
- Calibration of the number of colonies obtained at different optical densities.
- Transformation of competent E.coli (strain K12Z1) cells with constructs.
- Induction of the construct pLac cfp
E.coli cells transformed with the construct pLac cfp were cultured for 10 hours in Luria Bertani medium containing 50mg/ml Ampicillin.
The cells were then transferred into a minimal medium (M9 medium) and were induced with the following concentrations of IPTG. A stock concentration of 100mM was used.
| Sample | 100mM IPTG (ul) | 1mM IPTG (ul) | Inoculum volume from LB (ul) | Volume of M9 added (ml) |
|---|---|---|---|---|
| pLac cfp (0 uM) | - | - | 3 ul | 3 ml |
| pLac cfp (1 uM) | - | 3 ul | 3 ul | 3 ml |
| pLac cfp (10 uM) | - | 30 ul | 3 ul | 3 ml |
| pLac cfp (50 uM) | 1.5 ul | - | 3 ul | 3 ml |
| pLac cfp (100 uM) | 3 ul | - | 3 ul | 3 ml |
| pLac cfp (1000 uM) | 30 ul | - | 3 ul | 3 ml |
| K12Z1 (Autoflourescence) | - | - | 3 ul | 3 ml |
Cells were allowed to grow till the optical density was in the range of 0.05-0.1 (early exponential phase) and were imaged using a phase contrast microscope.
Results: The mean fluorescence of CFP was seen to increase with increasing concentrations of the IPTG.
- Induction of the construct pLac yfp
E.coli cells transformed with the construct pLac yfp were cultured for 10 hours in Luria Bertani medium containing 50mg/ml Ampicillin.
The cells were then transferred into a minimal medium (M9 medium) and were induced with the following concentrations of IPTG. A stock concentration of 100mM was used.
| Sample | 100mM IPTG (ul) | 1mM IPTG (ul) | Inoculum volume from LB (ul) | Volume of M9 added (ml) |
|---|---|---|---|---|
| pLac yfp (0 uM) | - | - | 3 ul | 3 ml |
| pLac yfp (1 uM) | - | 3 ul | 3 ul | 3 ml |
| pLac yfp (10 uM) | - | 30 ul | 3 ul | 3 ml |
| pLac yfp (50 uM) | 1.5 ul | - | 3 ul | 3 ml |
| pLac yfp (100 uM) | 3 ul | - | 3 ul | 3 ml |
| pLac yfp (1000 uM) | 30 ul | - | 3 ul | 3 ml |
| K12Z1 (Autoflourescence) | - | - | 3 ul | 3 ml |
Results: The mean fluorescence of YFP was seen to increase with increasing concentrations of the IPTG.
