"
Virginia Commonwealth/24 June 2009
From 2009.igem.org
(Difference between revisions)
(→Tasks) |
(→Tasks) |
||
| Line 44: | Line 44: | ||
===Tasks=== | ===Tasks=== | ||
---- | ---- | ||
| - | *1 | + | *1 Miniprep needs to be done on regrown backbone stock. |
| - | *2 | + | *2 Re-digest PSB3K3 and ligate the parts using the already digested promoter (J23102) and gene(EO0240). |
| + | *3 Run a gel electrophoresis on the digested PSB3K3. | ||
====Wetlab==== | ====Wetlab==== | ||
| - | *1 | + | *1 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.) |
| - | *2 | + | ** 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. | ||
| + | |||
| + | *2 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. | ||
| + | |||
| + | *3 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). | ||
| + | |||
| + | *4 Gel Electrophoresis was run, however no PSB3K3 was found on the plate. (details on page 12 of the iGEM notebook) | ||
| + | |||
| + | *5 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.) | ||
| + | |||
| + | [[User:MandM|MandM]] 02:22, 25 June 2009 (UTC) | ||
Revision as of 02:22, 25 June 2009
|
|
|
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Contents |
Wednesday 24 June 2009
Results
- 1
- 2
Tasks
- 1 Miniprep needs to be done on regrown backbone stock.
- 2 Re-digest PSB3K3 and ligate the parts using the already digested promoter (J23102) and gene(EO0240).
- 3 Run a gel electrophoresis on the digested PSB3K3.
Wetlab
- 1 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.
- 2 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:
- 3 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:
- 4 Gel Electrophoresis was run, however no PSB3K3 was found on the plate. (details on page 12 of the iGEM notebook)
- 5 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)
