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Virginia Commonwealth/24 June 2009
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==Wednesday 24 June 2009== | ==Wednesday 24 June 2009== | ||
===Results=== | ===Results=== | ||
| - | * | + | * Gel Electrophoresis was run on pSB3K3 only |
| - | * | + | * No DNA was visible |
| + | [[Image:2009-06-24 19digest02(psb3k3only).jpg]] | ||
| + | |||
| + | [[User:Trentay|Trentay]] 19:16, 5 August 2009 (UTC) | ||
---- | ---- | ||
| + | |||
===Tasks=== | ===Tasks=== | ||
| - | |||
*Miniprep needs to be done on regrown backbone stock. | *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). | *Re-digest PSB3K3 and ligate the parts using the already digested promoter (J23102) and gene(EO0240). | ||
*Run a gel electrophoresis on the digested PSB3K3. | *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.) | *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.) | ||
| Line 52: | Line 56: | ||
*** 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 | + | *Digestion was done on the PSB3K3 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. | ||
Latest revision as of 19:16, 5 August 2009
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Contents |
Wednesday 24 June 2009
Results
- Gel Electrophoresis was run on pSB3K3 only
- No DNA was visible
.jpg)
Trentay 19:16, 5 August 2009 (UTC)
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 PSB3K3 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)
