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Team:DTU-Denmark/Notebook/10 June 2013
From 2013.igem.org
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| + | {{:Team:DTU-Denmark/Templates/StartPage|10 June 2013}} | ||
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=208 lab= | =208 lab= | ||
| + | <hr/> | ||
== Main purposes today == | == Main purposes today == | ||
| + | <hr/> | ||
*helloworld-project | *helloworld-project | ||
*OD meassurements | *OD meassurements | ||
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==Who was in the lab== | ==Who was in the lab== | ||
| + | <hr/> | ||
Kristian, Jakob, Henrike | Kristian, Jakob, Henrike | ||
==Procedure== | ==Procedure== | ||
| + | <hr/> | ||
===OD measurements=== | ===OD measurements=== | ||
| + | <hr/> | ||
{| | {| | ||
!time!!OD600 measurement | !time!!OD600 measurement | ||
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===Making competent cells=== | ===Making competent cells=== | ||
| + | <hr/> | ||
Using the [[Team:DTU-Denmark/Methods/Competent Cells|Protocol]], we got from Andreas with following modifications to protocol: | Using the [[Team:DTU-Denmark/Methods/Competent Cells|Protocol]], we got from Andreas with following modifications to protocol: | ||
* made only half the amount | * made only half the amount | ||
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The competent cells were stored in -80 C freezer on the lower shelf | The competent cells were stored in -80 C freezer on the lower shelf | ||
| - | === | + | ===Autoclaving=== |
| + | <hr/> | ||
following was [[Team:DTU-Denmark/Methods/Autoclaving|autoclaved]]: | following was [[Team:DTU-Denmark/Methods/Autoclaving|autoclaved]]: | ||
*Eppendorf tubes | *Eppendorf tubes | ||
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===Solutions made=== | ===Solutions made=== | ||
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*0.1 M CaCl_2 and 15% glycerol from 10ml 1M CaCl_2 | *0.1 M CaCl_2 and 15% glycerol from 10ml 1M CaCl_2 | ||
*30ml 50% glycerol 70ml dm water | *30ml 50% glycerol 70ml dm water | ||
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*chlorampenicol stock 1ml with cencentration 50mg/ml | *chlorampenicol stock 1ml with cencentration 50mg/ml | ||
| - | == | + | ==Plates== |
| + | <hr/> | ||
*24 ps with 10ug/ml chlorampenicol (CAP) 5 mg to 500ml | *24 ps with 10ug/ml chlorampenicol (CAP) 5 mg to 500ml | ||
*33 ps with 30ug/ml Kanamycin 0.3ml of 30ug/ml to 500ml | *33 ps with 30ug/ml Kanamycin 0.3ml of 30ug/ml to 500ml | ||
| - | == | + | ==Transformation== |
| + | <hr/> | ||
Following the [http://parts.igem.org/Help:Protocols/Transformation official iGEM protocol] : | Following the [http://parts.igem.org/Help:Protocols/Transformation official iGEM protocol] : | ||
*BBa-I0500 plate 5 well 14N, then moved to -80C | *BBa-I0500 plate 5 well 14N, then moved to -80C | ||
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all 4 had 2 hours incubation. | all 4 had 2 hours incubation. | ||
| + | == Conclusion from today == | ||
| + | <hr/> | ||
| - | + | {{:Team:DTU-Denmark/Templates/EndPage}} | |
| - | + | ||
| - | + | ||
Revision as of 09:20, 1 July 2013
10 June 2013
Contents |
208 lab
Main purposes today
- helloworld-project
- OD meassurements
- autoclaving
- make competent cells
Who was in the lab
Kristian, Jakob, Henrike
Procedure
OD measurements
| time | OD600 measurement |
|---|---|
| 0.190 | |
| 11:16 | 0.289 |
| 11:28 | 0.356 |
| 11:32 | 0.390 |
| taken out 11:34 | 0.397 |
placed on ice
Making competent cells
Using the Protocol, we got from Andreas with following modifications to protocol:
- made only half the amount
- used falcon tubes
The competent cells were stored in -80 C freezer on the lower shelf
Autoclaving
following was autoclaved:
- Eppendorf tubes
- 3 x 100ml dm water
- 2 x 100ml 0.1M CaCl_2 and 15% glycerol
- 1 L bottle of 0.1M CaCl_2
- 1 L bottle of 50% glycerol
- 200ml bottle of 1M CaCl_2
Solutions made
- 0.1 M CaCl_2 and 15% glycerol from 10ml 1M CaCl_2
- 30ml 50% glycerol 70ml dm water
- 0.1 M CaCl_2 from 100ml 1M CaCl_2 and 900ml dm water
- chlorampenicol stock 1ml with cencentration 50mg/ml
Plates
- 24 ps with 10ug/ml chlorampenicol (CAP) 5 mg to 500ml
- 33 ps with 30ug/ml Kanamycin 0.3ml of 30ug/ml to 500ml
Transformation
Following the [http://parts.igem.org/Help:Protocols/Transformation official iGEM protocol] :
- BBa-I0500 plate 5 well 14N, then moved to -80C
- BBa-I14032 plate 3 well 18H, then moved to -80C
Transformation efficiency test (0.5,5,10,20,50)
Incubation 2 hours with 300 rpm
Plates:
- I0500 - no incubation on Kanamycin plate, one 20ul and one 200 ul
- I14032 - no incubation on CAP, one 20ul and one 200 ul
all 4 had 2 hours incubation.
Conclusion from today
