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| | + | |
| | + | {| align="center" |
| | + | |-valign="top" |
| | + | |align="center" width="150pt"|{{#calendar: title=Virginia_Commonwealth |year=2009 | month=05}} |
| | + | |align="center" width="150pt"|{{#calendar: title=Virginia_Commonwealth |year=2009 | month=06}} |
| | + | |align="center" width="150pt"|{{#calendar: title=Virginia_Commonwealth |year=2009 | month=07}} |
| | + | |align="center" width="150pt"|{{#calendar: title=Virginia_Commonwealth |year=2009 | month=08}} |
| | + | |align="center" width="150pt"|{{#calendar: title=Virginia_Commonwealth |year=2009 | month=09}} |
| | + | |align="center" width="150pt"|{{#calendar: title=Virginia_Commonwealth |year=2009 | month=10}} |
| | + | |} |
| | ==Tuesday 4 August 2009== | | ==Tuesday 4 August 2009== |
| | ===Results=== | | ===Results=== |
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| | * Possible inefficiency of BioBrick enzymes may have come from repeated freezing and thawing | | * Possible inefficiency of BioBrick enzymes may have come from repeated freezing and thawing |
| | ** They will be aliquoted into smaller amounts | | ** They will be aliquoted into smaller amounts |
| | + | * Gel Electrophoresis Results |
| | + | [[Image:2009-08-04 DigestI1352andpSB4C5.jpg|350px|thumb|none| From Left to Right: 100bp Ladder, Bioline Hyperladder, I1352(Digest), I1352(Miniprep), pSB4C5(Digest), pSB4C5(Miniprep)]] |
| | + | |
| | [[User:Trentay|Trentay]] 22:25, 4 August 2009 (UTC) | | [[User:Trentay|Trentay]] 22:25, 4 August 2009 (UTC) |
| | ---- | | ---- |
| | + | |
| | ===Tasks=== | | ===Tasks=== |
| | ''Maria and Afton'' | | ''Maria and Afton'' |
| Line 20: |
Line 62: |
| | ====Wetlab==== | | ====Wetlab==== |
| | * New BioBrick Assembly Kit came in and the following were aliquoted into their respective amounts | | * New BioBrick Assembly Kit came in and the following were aliquoted into their respective amounts |
| - | ** XbaI (10 µL)
| + | {|border="1" |
| - | ** SpeI (5 µL)
| + | |+ |
| - | ** PstI (10 µL)
| + | |- |
| - | ** EcoRI-HF (10 µL)
| + | ! Name !! Amount/Tube !! # Tubes !! Extra in Stock !! |
| - | ** BSA (5 µL)
| + | |- |
| - | ** T4 Ligase Buffer (10 µL)
| + | ! XbaI |
| - | ** T4 Ligase (5 µL)
| + | | 10 µL || 15 || No || |
| - | [[User:Trentay|Trentay]] 22:33, 4 August 2009 (UTC) | + | |- |
| | + | ! SpeI |
| | + | | 5 µL || 10 || No || |
| | + | |- |
| | + | ! PstI |
| | + | | 10 µL || 25 || Yes || |
| | + | |- |
| | + | ! EcoRI-HF |
| | + | | 10 µL || 25 || Yes || |
| | + | |- |
| | + | ! BSA |
| | + | | 5 µL || 8 || Yes || |
| | + | |- |
| | + | ! T4 Ligase Buffer |
| | + | | 10 µL || 25 || Yes || |
| | + | |- |
| | + | ! T4 Ligase |
| | + | | 10 µL || 8 || No || |
| | + | |} |
| | + | * To show how many times a tube has been thawed and refrozen, a hash mark will be placed on the side of the tube, each time it is thawed as a reference for others to see |
| | + | [[User:Trentay|Trentay]] 15:24, 5 August 2009 (UTC) |
| | | | |
| | ''Maria and Afton'' | | ''Maria and Afton'' |
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| | * pSB1C3 w/ P1010 was removed from Well 5E on Plate 1 of the Distribution Kit and resuspended in 10µL of TE Buffer. | | * pSB1C3 w/ P1010 was removed from Well 5E on Plate 1 of the Distribution Kit and resuspended in 10µL of TE Buffer. |
| | * Transformation was done using DB 3.1 | | * Transformation was done using DB 3.1 |
| - | {| cellpadding="10" align="center" border="0.9" padding="5" | + | {|border="1" |
| | |+ | | |+ |
| | |- | | |- |
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| | |- | | |- |
| | ! pSB1C3 | | ! pSB1C3 |
| - | | CM|| || | + | | CM|| n/a || |
| | |- | | |- |
| | ! (+) Control | | ! (+) Control |
| - | | LB || || | + | | LB || n/a || |
| | |- | | |- |
| | ! (+) Control zapped | | ! (+) Control zapped |
| - | | LB || || | + | | LB || n/a || |
| | |- | | |- |
| | ! (-) Control | | ! (-) Control |
| - | | CM || || | + | | CM || n/a || |
| | |} | | |} |
| | [[User:Trentay|Trentay]] 22:33, 4 August 2009 (UTC) | | [[User:Trentay|Trentay]] 22:33, 4 August 2009 (UTC) |
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| [http://2009.igem.org/Virginia_Commonwealth/30_May_2009 30]
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| [http://2009.igem.org/Virginia_Commonwealth/1_June_2009 1]
| [http://2009.igem.org/Virginia_Commonwealth/2_June_2009 2]
| [http://2009.igem.org/Virginia_Commonwealth/3_June_2009 3]
| [http://2009.igem.org/Virginia_Commonwealth/4_June_2009 4]
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| [http://2009.igem.org/Virginia_Commonwealth/6_August_2009 6]
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| [http://2009.igem.org/Virginia_Commonwealth/13_August_2009 13]
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| [http://2009.igem.org/Virginia_Commonwealth/3_September_2009 3]
| [http://2009.igem.org/Virginia_Commonwealth/4_September_2009 4]
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| [http://2009.igem.org/Virginia_Commonwealth/10_September_2009 10]
| [http://2009.igem.org/Virginia_Commonwealth/11_September_2009 11]
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| [http://2009.igem.org/Virginia_Commonwealth/1_October_2009 1]
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| [http://2009.igem.org/Virginia_Commonwealth/8_October_2009 8]
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| [http://2009.igem.org/Virginia_Commonwealth/29_October_2009 29]
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Tuesday 4 August 2009
Results
Maria and Afton
- Overnight culture of I1352 plated on AMP:
- Sample taken from plate after PSB4C5 ligation attempt successfully grew expressing RFP. The color of the sample was pink
- Sample taken from plate after ligation with itself did not grow at all, and expressed no RFP
- There is a possibility the pSB4C5 backbone may be a bad part
- previous years it has not functioned properly
- Gel electrophoresis indicates DNA Gel Purification may be necessary
- Possible inefficiency of BioBrick enzymes may have come from repeated freezing and thawing
- They will be aliquoted into smaller amounts
- Gel Electrophoresis Results
From Left to Right: 100bp Ladder, Bioline Hyperladder, I1352(Digest), I1352(Miniprep), pSB4C5(Digest), pSB4C5(Miniprep)
Trentay 22:25, 4 August 2009 (UTC)
Tasks
Maria and Afton
- Run a Gel Electrophoresis
- Transform and grow up pSB1C3 with the death gene P1010
Trentay 22:11, 4 August 2009 (UTC)
Wetlab
- New BioBrick Assembly Kit came in and the following were aliquoted into their respective amounts
| Name | Amount/Tube | # Tubes | Extra in Stock |
|
| XbaI
| 10 µL | 15 | No |
|
| SpeI
| 5 µL | 10 | No |
|
| PstI
| 10 µL | 25 | Yes |
|
| EcoRI-HF
| 10 µL | 25 | Yes |
|
| BSA
| 5 µL | 8 | Yes |
|
| T4 Ligase Buffer
| 10 µL | 25 | Yes |
|
| T4 Ligase
| 10 µL | 8 | No |
|
- To show how many times a tube has been thawed and refrozen, a hash mark will be placed on the side of the tube, each time it is thawed as a reference for others to see
Trentay 15:24, 5 August 2009 (UTC)
Maria and Afton
- Gel Electrophoresis was run
- pSB4C5 and I1352 digested and undigested (right after Miniprep) were run on a gel and results showed excessive bands in the undigested DNA. DNA Gel Purification will be done on future Miniprepped DNA
- pSB1C3 w/ P1010 was removed from Well 5E on Plate 1 of the Distribution Kit and resuspended in 10µL of TE Buffer.
- Transformation was done using DB 3.1
| Name | Plate | Growth Observation |
|
| pSB1C3
| CM | n/a |
|
| (+) Control
| LB | n/a |
|
| (+) Control zapped
| LB | n/a |
|
| (-) Control
| CM | n/a |
|
Trentay 22:33, 4 August 2009 (UTC)
|
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