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Virginia Commonwealth/24 June 2009
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===Tasks=== | ===Tasks=== | ||
---- | ---- | ||
| - | * | + | *Miniprep needs to be done on regrown backbone stock. |
| - | * | + | *Re-digest PSB3K3 and ligate the parts using the already digested promoter (J23102) and gene(EO0240). |
| - | * | + | *Run a gel electrophoresis on the digested PSB3K3. |
====Wetlab==== | ====Wetlab==== | ||
| - | * | + | *The promoter DNA from the mini-preps run by Afton on the 12th were Concentrated using the "Ethanol precipitation of nucleic acids" protocol on OpenWetWare.(Detailed notes can be found on page 10 of the iGEM notebook.) |
** Note: A spec needs to be run on the concentrated DNA to obtain the current concentrations and volumes. | ** Note: A spec needs to be run on the concentrated DNA to obtain the current concentrations and volumes. | ||
*** This should be held off until the backbone problems are rectified as the DNA may degrade over time. | *** This should be held off until the backbone problems are rectified as the DNA may degrade over time. | ||
| - | * | + | *Digestion was done on the PSB3K# using the enzymes ECO RI-HF and PstI, in the respective order. |
**Problems: | **Problems: | ||
*** The 80 deg. Celsius bath was not ready during the incubation so the cells were kept in the incubator longer than 15 minutes. | *** The 80 deg. Celsius bath was not ready during the incubation so the cells were kept in the incubator longer than 15 minutes. | ||
| - | * | + | *Ligation was performed along with transformation and is growing up overnight. The results will be listed above in the morning. |
**Problems: | **Problems: | ||
***Maria was distracted while transforming and used 5 increments of 200 uL of the SOC because the 1000 uL pipet was not ready. The delay may have caused decreased efficiency (although it was not more than 1 minute before the 1st 200uL). | ***Maria was distracted while transforming and used 5 increments of 200 uL of the SOC because the 1000 uL pipet was not ready. The delay may have caused decreased efficiency (although it was not more than 1 minute before the 1st 200uL). | ||
| - | * | + | *Gel Electrophoresis was run, however no PSB3K3 was found on the plate. (details on page 12 of the iGEM notebook) |
| - | * | + | *PSB3K3 was re-plated using the plasmid from the Biobrick well stock. Colonies should be picked tomorrow afternoon. |
(All data is on pages 10-12 in the iGEM lab notebook for more detail.) | (All data is on pages 10-12 in the iGEM lab notebook for more detail.) | ||
[[User:MandM|MandM]] 02:22, 25 June 2009 (UTC) | [[User:MandM|MandM]] 02:22, 25 June 2009 (UTC) | ||
Revision as of 15:09, 25 June 2009
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Contents |
Wednesday 24 June 2009
Results
- 1
- 2
Tasks
- Miniprep needs to be done on regrown backbone stock.
- Re-digest PSB3K3 and ligate the parts using the already digested promoter (J23102) and gene(EO0240).
- Run a gel electrophoresis on the digested PSB3K3.
Wetlab
- The promoter DNA from the mini-preps run by Afton on the 12th were Concentrated using the "Ethanol precipitation of nucleic acids" protocol on OpenWetWare.(Detailed notes can be found on page 10 of the iGEM notebook.)
- Note: A spec needs to be run on the concentrated DNA to obtain the current concentrations and volumes.
- This should be held off until the backbone problems are rectified as the DNA may degrade over time.
- Note: A spec needs to be run on the concentrated DNA to obtain the current concentrations and volumes.
- Digestion was done on the PSB3K# using the enzymes ECO RI-HF and PstI, in the respective order.
- Problems:
- The 80 deg. Celsius bath was not ready during the incubation so the cells were kept in the incubator longer than 15 minutes.
- Problems:
- Ligation was performed along with transformation and is growing up overnight. The results will be listed above in the morning.
- Problems:
- Maria was distracted while transforming and used 5 increments of 200 uL of the SOC because the 1000 uL pipet was not ready. The delay may have caused decreased efficiency (although it was not more than 1 minute before the 1st 200uL).
- Problems:
- Gel Electrophoresis was run, however no PSB3K3 was found on the plate. (details on page 12 of the iGEM notebook)
- PSB3K3 was re-plated using the plasmid from the Biobrick well stock. Colonies should be picked tomorrow afternoon.
(All data is on pages 10-12 in the iGEM lab notebook for more detail.)
MandM 02:22, 25 June 2009 (UTC)
