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| | ==Tuesday 11 August 2009== | | ==Tuesday 11 August 2009== |
| | ===Results=== | | ===Results=== |
| | + | ''Kevin and Adam'' |
| | + | *I have cell growth on all plates. Yesterday's work was successful. Colonies number from 2-5 colonies per plate at 4:00 PM. I will wait until later to pick a colony. |
| | + | |
| | ''Maria and Afton'' | | ''Maria and Afton'' |
| | * 1kb ladder has not come in yet so, digests and minipreps will be run on gels when it comes in | | * 1kb ladder has not come in yet so, digests and minipreps will be run on gels when it comes in |
| | * Plated colonies from Friday (8/7/09) transformations are strongly espressing RFP | | * Plated colonies from Friday (8/7/09) transformations are strongly espressing RFP |
| | ** It might be better to pick colonies 3 to 4 days after transformation when colonies are strongly espressing RFP, instead of when the e. coli are pink the day after transformation | | ** It might be better to pick colonies 3 to 4 days after transformation when colonies are strongly espressing RFP, instead of when the e. coli are pink the day after transformation |
| | + | *Spectrophotometry Data |
| | + | {| cellpadding="5" border="1" padding="5" |
| | + | |+ |
| | + | |- |
| | + | ! Part No.!! A260 (nm) !! A280 (nm) !! A260/A280 !! Vol. (µL) !! Conc. (µg/mL) !! Amt (µg) |
| | + | |- |
| | + | | pSB1C3 p1010 || 0.012 || 0.008 || 1.5 || 54 || 24 || 1.3 |
| | + | |- |
| | + | | J23100 || 0.029 || 0.023 || 1.26 || 35 || 58 || 2.03 |
| | + | |- |
| | + | | J23104 || 0.030 || 0.026 || 1.15 || 38 || 60 || 2.28 |
| | + | |- |
| | + | | J23102 || 0.022 || 0.019 || 1.16 || 40 || 44 || 1.76 |
| | + | |- |
| | + | | J23103 || 0.021 || 0.015 || 1.40 || 45 || 42 || 1.89 |
| | + | |- |
| | + | | J06702 || 0.029 || 0.018 || 1.61 || 117 || 58 || 6.79 |
| | + | |} |
| | + | [[User:Trentay|Trentay]] 19:17, 11 August 2009 (UTC) |
| | ---- | | ---- |
| | + | |
| | ===Tasks=== | | ===Tasks=== |
| | ''Maria and Afton'' | | ''Maria and Afton'' |
| Line 52: |
Line 75: |
| | * Transform parts into NEB10β | | * Transform parts into NEB10β |
| | * Update Wiki | | * Update Wiki |
| - | * Make overnight cultures of I1352/I1352, I1352/pSB1C3 | + | * Make overnight cultures of I1352/pSB1C3 |
| - | * Make overnight cultures of pSB1C3 p1010 from cryogenic storage to Miniprep tomorrow | + | ** I1352/I1352 will not be kept because it never expressed RFP and there are plenty of stock of I1352 with Amp resistance |
| | + | * Make overnight cultures of pSB1C3 p1010 from cryogenic storage to Miniprep tomorrow |
| | + | * Make more LB+Cm plates |
| | [[User:Trentay|Trentay]] 14:43, 11 August 2009 (UTC) | | [[User:Trentay|Trentay]] 14:43, 11 August 2009 (UTC) |
| | ---- | | ---- |
| Line 60: |
Line 85: |
| | * Spectrophotometry was done | | * Spectrophotometry was done |
| | * Digest was done | | * Digest was done |
| - | [[User:Trentay|Trentay]] 14:43, 11 August 2009 (UTC) | + | * Ligation was done |
| | + | * Transformation was done |
| | + | * Overnight cultures were made |
| | + | * LB+Cm plates were made |
| | + | [[User:Trentay|Trentay]] 20:12, 11 August 2009 (UTC) |
| | + | |
| | + | ''Adam and Kevin'' |
| | + | * The growth on the plates remains slow. No colonies show fluorescence yet, for the sake of better selection I am going to wait until early tomorrow morning to pick colonies. I will cultivate them tomorrow and then store them in the evening. Hopefully we will be able to measure transcription levels Thursday. |
| | + | -[[User:Bussingkm|Bussingkm]] 21:16, 11 August 2009 (UTC) |
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| [http://2009.igem.org/Virginia_Commonwealth/21_May_2009 21]
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| [http://2009.igem.org/Virginia_Commonwealth/27_May_2009 27]
| [http://2009.igem.org/Virginia_Commonwealth/28_May_2009 28]
| [http://2009.igem.org/Virginia_Commonwealth/29_May_2009 29]
| [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]
| [http://2009.igem.org/Virginia_Commonwealth/5_June_2009 5]
| [http://2009.igem.org/Virginia_Commonwealth/6_June_2009 6]
| [http://2009.igem.org/Virginia_Commonwealth/7_June_2009 7]
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| [http://2009.igem.org/Virginia_Commonwealth/9_June_2009 9]
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| [http://2009.igem.org/Virginia_Commonwealth/11_June_2009 11]
| [http://2009.igem.org/Virginia_Commonwealth/12_June_2009 12]
| [http://2009.igem.org/Virginia_Commonwealth/13_June_2009 13]
| [http://2009.igem.org/Virginia_Commonwealth/14_June_2009 14]
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| [http://2009.igem.org/Virginia_Commonwealth/16_June_2009 16]
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| [http://2009.igem.org/Virginia_Commonwealth/28_June_2009 28]
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| [http://2009.igem.org/Virginia_Commonwealth/3_July_2009 3]
| [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/4_July_2009&action=edit 4]
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| [http://2009.igem.org/Virginia_Commonwealth/7_July_2009 7]
| [http://2009.igem.org/Virginia_Commonwealth/8_July_2009 8]
| [http://2009.igem.org/Virginia_Commonwealth/9_July_2009 9]
| [http://2009.igem.org/Virginia_Commonwealth/10_July_2009 10]
| [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/11_July_2009&action=edit 11]
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| [http://2009.igem.org/Virginia_Commonwealth/15_July_2009 15]
| [http://2009.igem.org/Virginia_Commonwealth/16_July_2009 16]
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| [http://2009.igem.org/Virginia_Commonwealth/23_July_2009 23]
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| [http://2009.igem.org/Virginia_Commonwealth/30_July_2009 30]
| [http://2009.igem.org/Virginia_Commonwealth/31_July_2009 31]
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| [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/2_August_2009&action=edit 2]
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| [http://2009.igem.org/Virginia_Commonwealth/4_August_2009 4]
| [http://2009.igem.org/Virginia_Commonwealth/5_August_2009 5]
| [http://2009.igem.org/Virginia_Commonwealth/6_August_2009 6]
| [http://2009.igem.org/Virginia_Commonwealth/7_August_2009 7]
| [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/8_August_2009&action=edit 8]
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| [http://2009.igem.org/Virginia_Commonwealth/11_August_2009 11]
| [http://2009.igem.org/Virginia_Commonwealth/12_August_2009 12]
| [http://2009.igem.org/Virginia_Commonwealth/13_August_2009 13]
| [http://2009.igem.org/Virginia_Commonwealth/14_August_2009 14]
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| [http://2009.igem.org/Virginia_Commonwealth/20_August_2009 20]
| [http://2009.igem.org/Virginia_Commonwealth/21_August_2009 21]
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| [http://2009.igem.org/Virginia_Commonwealth/27_August_2009 27]
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| [http://2009.igem.org/Virginia_Commonwealth/1_September_2009 1]
| [http://2009.igem.org/Virginia_Commonwealth/2_September_2009 2]
| [http://2009.igem.org/Virginia_Commonwealth/3_September_2009 3]
| [http://2009.igem.org/Virginia_Commonwealth/4_September_2009 4]
| [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/5_September_2009&action=edit 5]
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| [http://2009.igem.org/Virginia_Commonwealth/8_September_2009 8]
| [http://2009.igem.org/Virginia_Commonwealth/9_September_2009 9]
| [http://2009.igem.org/Virginia_Commonwealth/10_September_2009 10]
| [http://2009.igem.org/Virginia_Commonwealth/11_September_2009 11]
| [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/12_September_2009&action=edit 12]
| [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/13_September_2009&action=edit 13]
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| [http://2009.igem.org/Virginia_Commonwealth/15_September_2009 15]
| [http://2009.igem.org/Virginia_Commonwealth/16_September_2009 16]
| [http://2009.igem.org/Virginia_Commonwealth/17_September_2009 17]
| [http://2009.igem.org/Virginia_Commonwealth/18_September_2009 18]
| [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/19_September_2009&action=edit 19]
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| [http://2009.igem.org/Virginia_Commonwealth/22_September_2009 22]
| [http://2009.igem.org/Virginia_Commonwealth/23_September_2009 23]
| [http://2009.igem.org/Virginia_Commonwealth/24_September_2009 24]
| [http://2009.igem.org/Virginia_Commonwealth/25_September_2009 25]
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| [http://2009.igem.org/Virginia_Commonwealth/1_October_2009 1]
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| [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/3_October_2009&action=edit 3]
| [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/4_October_2009&action=edit 4]
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| [http://2009.igem.org/Virginia_Commonwealth/6_October_2009 6]
| [http://2009.igem.org/Virginia_Commonwealth/7_October_2009 7]
| [http://2009.igem.org/Virginia_Commonwealth/8_October_2009 8]
| [http://2009.igem.org/Virginia_Commonwealth/9_October_2009 9]
| [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/10_October_2009&action=edit 10]
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| [http://2009.igem.org/Virginia_Commonwealth/14_October_2009 14]
| [http://2009.igem.org/Virginia_Commonwealth/15_October_2009 15]
| [http://2009.igem.org/Virginia_Commonwealth/16_October_2009 16]
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| [http://2009.igem.org/Virginia_Commonwealth/18_October_2009 18]
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| [http://2009.igem.org/Virginia_Commonwealth/20_October_2009 20]
| [http://2009.igem.org/Virginia_Commonwealth/21_October_2009 21]
| [http://2009.igem.org/Virginia_Commonwealth/22_October_2009 22]
| [http://2009.igem.org/Virginia_Commonwealth/23_October_2009 23]
| [http://2009.igem.org/Virginia_Commonwealth/24_October_2009 24]
| [http://2009.igem.org/Virginia_Commonwealth/25_October_2009 25]
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| [http://2009.igem.org/Virginia_Commonwealth/27_October_2009 27]
| [http://2009.igem.org/Virginia_Commonwealth/28_October_2009 28]
| [http://2009.igem.org/Virginia_Commonwealth/29_October_2009 29]
| [http://2009.igem.org/Virginia_Commonwealth/30_October_2009 30]
| [http://2009.igem.org/wiki/index.php?title=Virginia_Commonwealth/31_October_2009&action=edit 31]
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Tuesday 11 August 2009
Results
Kevin and Adam
- I have cell growth on all plates. Yesterday's work was successful. Colonies number from 2-5 colonies per plate at 4:00 PM. I will wait until later to pick a colony.
Maria and Afton
- 1kb ladder has not come in yet so, digests and minipreps will be run on gels when it comes in
- Plated colonies from Friday (8/7/09) transformations are strongly espressing RFP
- It might be better to pick colonies 3 to 4 days after transformation when colonies are strongly espressing RFP, instead of when the e. coli are pink the day after transformation
- Spectrophotometry Data
| Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg)
|
| pSB1C3 p1010 | 0.012 | 0.008 | 1.5 | 54 | 24 | 1.3
|
| J23100 | 0.029 | 0.023 | 1.26 | 35 | 58 | 2.03
|
| J23104 | 0.030 | 0.026 | 1.15 | 38 | 60 | 2.28
|
| J23102 | 0.022 | 0.019 | 1.16 | 40 | 44 | 1.76
|
| J23103 | 0.021 | 0.015 | 1.40 | 45 | 42 | 1.89
|
| J06702 | 0.029 | 0.018 | 1.61 | 117 | 58 | 6.79
|
Trentay 19:17, 11 August 2009 (UTC)
Tasks
Maria and Afton
- Take Spectrophotometry measurements of pSB1C3 p1010, J06702, J23100, J23102, J23103, J23104
- Need to locate J23100 to be digested another day
- Digest above parts
- Ligate J06702 and pSB1C3 with the following promoters: 100, 102, 103, 104
- Transform parts into NEB10β
- Update Wiki
- Make overnight cultures of I1352/pSB1C3
- I1352/I1352 will not be kept because it never expressed RFP and there are plenty of stock of I1352 with Amp resistance
- Make overnight cultures of pSB1C3 p1010 from cryogenic storage to Miniprep tomorrow
- Make more LB+Cm plates
Trentay 14:43, 11 August 2009 (UTC)
Wetlab
Maria and Afton
- Spectrophotometry was done
- Digest was done
- Ligation was done
- Transformation was done
- Overnight cultures were made
- LB+Cm plates were made
Trentay 20:12, 11 August 2009 (UTC)
Adam and Kevin
- The growth on the plates remains slow. No colonies show fluorescence yet, for the sake of better selection I am going to wait until early tomorrow morning to pick colonies. I will cultivate them tomorrow and then store them in the evening. Hopefully we will be able to measure transcription levels Thursday.
-Bussingkm 21:16, 11 August 2009 (UTC)
|
"
