E-Notebook
From 2007.igem.org
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#Calibration of the number of colonies obtained at different optical densities. | #Calibration of the number of colonies obtained at different optical densities. | ||
#Transformation of competent E.coli (strain K12Z1) cells with constructs. | #Transformation of competent E.coli (strain K12Z1) cells with constructs. | ||
| - | #Induction of the construct pLac cfp | + | #'''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. | E.coli cells transformed with the construct pLac cfp were cultured for 10 hours in Luria Bertani medium containing 50mg/ml Ampicillin. | ||
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|pLac cfp (1 uM) | |pLac cfp (1 uM) | ||
| - | | - | ||
| - | | | + | |3 ul |
|3 ul | |3 ul | ||
|3 ml | |3 ml | ||
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|pLac cfp (10 uM) | |pLac cfp (10 uM) | ||
| - | | - | ||
| - | | | + | |30 ul |
|3 ul | |3 ul | ||
|3 ml | |3 ml | ||
|-align="center" | |-align="center" | ||
|pLac cfp (50 uM) | |pLac cfp (50 uM) | ||
| - | | | + | |1.5 ul |
| - | | - | ||
|3 ul | |3 ul | ||
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|-align="center" | |-align="center" | ||
|pLac cfp (100 uM) | |pLac cfp (100 uM) | ||
| - | | | + | |3 ul |
| - | | - | ||
|3 ul | |3 ul | ||
| Line 303: | Line 303: | ||
|-align="center" | |-align="center" | ||
|pLac cfp (1000 uM) | |pLac cfp (1000 uM) | ||
| - | | | + | |30 ul |
| - | | - | ||
|3 ul | |3 ul | ||
|3 ml | |3 ml | ||
|-align="center" | |-align="center" | ||
| - | | | + | |K12Z1 (Autoflourescence) |
| - | | - | ||
| - | | - | ||
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|3 ml | |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. | ||
== The Week Ahead == | == The Week Ahead == | ||
Revision as of 07:00, 27 June 2007
| The official wiki of the NCBS iGEM 2007 Team |
| 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
|
Contents |
e-notebook
| May 07 | June 07 | July 07 | ||||||||||||||||||||||||||||||||
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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.
