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Virginia Commonwealth/10 July 2009
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(New page: ==Friday 10 July 2009== ===Results=== ''Afton and Maria'' *Overnight culture was successful ~~~~ ---- ===Tasks=== ''Afton and Maria'' * Make stocks of parts J23106 and J06702 * Miniprep o...) |
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==Friday 10 July 2009== | ==Friday 10 July 2009== | ||
===Results=== | ===Results=== | ||
''Afton and Maria'' | ''Afton and Maria'' | ||
*Overnight culture was successful | *Overnight culture was successful | ||
| + | * Miniprep/Spectrophotometry Results | ||
| + | {| cellpadding="5" border="1" padding="5" | ||
| + | |+ | ||
| + | |- | ||
| + | ! Part No.!! A260 (nm) !! A280 (nm) !! A260/A280 !! Vol. (µL) !! Conc. (µg/mL) !! Amt (µg) | ||
| + | |- | ||
| + | | J23106 || 0.015 || 0.004 || 3.750 || 120 || 22.5 || 2.700 | ||
| + | |- | ||
| + | | J06702 || 0.022 || 0.013 || 1.692 || 164 || 33.0 || 5.412 | ||
| + | |- | ||
| + | | J23100 || 0.036 || 0.025 || 1.440 || 54 || 22.5 || 2.808 | ||
| + | |} | ||
| + | |||
[[User:Trentay|Trentay]] 13:13, 10 July 2009 (UTC) | [[User:Trentay|Trentay]] 13:13, 10 July 2009 (UTC) | ||
| + | |||
| + | ''Craig and Clay'' | ||
| + | * The overnight culture of the three parts related to limonene synthesis were unsuccessful. Possible reasons include bad parts, errors in electrotransformation procedure, and incorrect resistance. | ||
---- | ---- | ||
| Line 10: | Line 64: | ||
* Make stocks of parts J23106 and J06702 | * Make stocks of parts J23106 and J06702 | ||
* Miniprep of parts J23106 and J06702 | * Miniprep of parts J23106 and J06702 | ||
| - | * Digestion of parts J23106 and J06702 | + | * Digestion of parts J23106 and J06702, (and J23100) |
* Electrophoresis test of parts J23106 and J06702 | * Electrophoresis test of parts J23106 and J06702 | ||
* May Ligate parts with backbones pSB1C3 and pSB4C5 and store DNA | * May Ligate parts with backbones pSB1C3 and pSB4C5 and store DNA | ||
| Line 17: | Line 71: | ||
** Write out a plan to organize ideas | ** Write out a plan to organize ideas | ||
[[User:Trentay|Trentay]] 13:13, 10 July 2009 (UTC) | [[User:Trentay|Trentay]] 13:13, 10 July 2009 (UTC) | ||
| + | |||
| + | [[User:Trentay|Trentay]] 17:05, 13 August 2009 (UTC) | ||
| + | |||
| + | ''Craig and Clay'' | ||
| + | * BBa_I742111, BBa_K118024, and BBa_K118025 need to be plated again. | ||
| + | * If the new plates also show no growth, electrotransformation of the parts will be performed again. | ||
| + | * If this proves to be unsuccessful, the cells from this second electrotransformation will also be plated a second time. The cells will also be plated on non-resistant media. | ||
| + | * If growth is again unsuccessful, the cells grown on the non-resistant media will be streaked on kanamycin and chloramphenicol plates. | ||
---- | ---- | ||
====Wetlab==== | ====Wetlab==== | ||
''Afton and Maria'' | ''Afton and Maria'' | ||
* Dilution of overnight culture | * Dilution of overnight culture | ||
| - | **2 mL of overnight culture was added to 4 mL of LB broth | + | **2 mL of overnight culture was added to 4 mL of LB broth (6 mL) |
***2 dilutions were done for each test tube | ***2 dilutions were done for each test tube | ||
**Cells not used in the MiniPrep will be stored in the -80 degree Celsius freezer | **Cells not used in the MiniPrep will be stored in the -80 degree Celsius freezer | ||
| - | [[User:Trentay|Trentay]] | + | * 500mL of 30 percent Glycerol stock was made |
| + | * Cell stocks were made | ||
| + | ** 10 vials of J06702 were put in the -80 degree Celsius freezer | ||
| + | ** 5 vials of J23106 were put in the -80 degree Celsius freezer | ||
| + | * MiniPrep was done on parts: J23106 and J6702 | ||
| + | **Amount of DNA was calculated with a Spectrophotometer | ||
| + | * Digestion was performed on MiniPrepped parts | ||
| + | **There are two digested J06702 PCR tubes | ||
| + | **Digests will be frozen until Monday | ||
| + | ***Electrophoresis test will be held off until then | ||
| + | ***Ligation as well | ||
| + | [[User:Trentay|Trentay]] 20:35, 10 July 2009 (UTC) | ||
| + | |||
| + | ''Craig and Clay'' | ||
| + | * The electrotransformed parts were plated again (ampicillin plates) due to unsuccessful growth. Fifty microliters was used instead of 35 microliters. | ||
Latest revision as of 17:13, 13 August 2009
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Contents |
Friday 10 July 2009
Results
Afton and Maria
- Overnight culture was successful
- Miniprep/Spectrophotometry Results
| Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg) |
|---|---|---|---|---|---|---|
| J23106 | 0.015 | 0.004 | 3.750 | 120 | 22.5 | 2.700 |
| J06702 | 0.022 | 0.013 | 1.692 | 164 | 33.0 | 5.412 |
| J23100 | 0.036 | 0.025 | 1.440 | 54 | 22.5 | 2.808 |
Trentay 13:13, 10 July 2009 (UTC)
Craig and Clay
- The overnight culture of the three parts related to limonene synthesis were unsuccessful. Possible reasons include bad parts, errors in electrotransformation procedure, and incorrect resistance.
Tasks
Afton and Maria
- Make stocks of parts J23106 and J06702
- Miniprep of parts J23106 and J06702
- Digestion of parts J23106 and J06702, (and J23100)
- Electrophoresis test of parts J23106 and J06702
- May Ligate parts with backbones pSB1C3 and pSB4C5 and store DNA
- Write out complete plan for experimentation
- Make plans to MaxiPrep
- Write out a plan to organize ideas
Trentay 13:13, 10 July 2009 (UTC)
Trentay 17:05, 13 August 2009 (UTC)
Craig and Clay
- BBa_I742111, BBa_K118024, and BBa_K118025 need to be plated again.
- If the new plates also show no growth, electrotransformation of the parts will be performed again.
- If this proves to be unsuccessful, the cells from this second electrotransformation will also be plated a second time. The cells will also be plated on non-resistant media.
- If growth is again unsuccessful, the cells grown on the non-resistant media will be streaked on kanamycin and chloramphenicol plates.
Wetlab
Afton and Maria
- Dilution of overnight culture
- 2 mL of overnight culture was added to 4 mL of LB broth (6 mL)
- 2 dilutions were done for each test tube
- Cells not used in the MiniPrep will be stored in the -80 degree Celsius freezer
- 2 mL of overnight culture was added to 4 mL of LB broth (6 mL)
- 500mL of 30 percent Glycerol stock was made
- Cell stocks were made
- 10 vials of J06702 were put in the -80 degree Celsius freezer
- 5 vials of J23106 were put in the -80 degree Celsius freezer
- MiniPrep was done on parts: J23106 and J6702
- Amount of DNA was calculated with a Spectrophotometer
- Digestion was performed on MiniPrepped parts
- There are two digested J06702 PCR tubes
- Digests will be frozen until Monday
- Electrophoresis test will be held off until then
- Ligation as well
Trentay 20:35, 10 July 2009 (UTC)
Craig and Clay
- The electrotransformed parts were plated again (ampicillin plates) due to unsuccessful growth. Fifty microliters was used instead of 35 microliters.
