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Virginia Commonwealth/7 July 2009
From 2009.igem.org
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===Results=== | ===Results=== | ||
* Everyone is still waiting on electrotransformation cuvettes to proceed with projects | * Everyone is still waiting on electrotransformation cuvettes to proceed with projects | ||
| - | '' | + | ''Maria and Afton'' |
| - | *pSB3K3 backbone did not show up on the gel | + | * Miniprep Results: |
| + | {| cellpadding="5" border="1" padding="5" | ||
| + | |+ | ||
| + | |- | ||
| + | ! Part No.!! A260 (nm) !! A280 (nm) !! A260/A280 !! Vol. (µL) !! Conc. (µg/mL) !! Amt (µg) | ||
| + | |- | ||
| + | | pSB3K3 (1) || 0.011 || 0.010 || 1.10 || 35 || 11 || 0.385 | ||
| + | |- | ||
| + | | pSB3K3 (2) || 0.010 || 0.008 || 1.25 || 35 || 10 || 0.350 | ||
| + | |- | ||
| + | | pSB3K3 (3) || 0.010 || 0.008 || 1.25 || 30 || 11 || 0.300 | ||
| + | |- | ||
| + | | pSB3K3 (All) || 0.010 || 0.010 || 1.00 || 97 || 20 || 1.940 | ||
| + | |} | ||
| + | * Individually the concentrations of pSB3K3 were too small for digestion, so all of the Miniprepped samples were combined and measurements were retaken. | ||
| + | |||
| + | *pSB3K3 backbone did not show up on the gel. Only the ladder is visible | ||
| + | [[image:2009-07-07 digest03 pSB3K3.jpg]] | ||
| + | |||
[[User:Trentay|Trentay]] 23:23, 7 July 2009 (UTC) | [[User:Trentay|Trentay]] 23:23, 7 July 2009 (UTC) | ||
---- | ---- | ||
| Line 47: | Line 65: | ||
===Tasks=== | ===Tasks=== | ||
* Continue research and writing paper drafts | * Continue research and writing paper drafts | ||
| - | '' | + | ''Maria and Afton'' |
* Take Spectrometer measurements to get DNA amounts of pSB3K3 | * Take Spectrometer measurements to get DNA amounts of pSB3K3 | ||
* Digestion of pSB3K3 | * Digestion of pSB3K3 | ||
| Line 53: | Line 71: | ||
* Run Electrophoresis test on digestion | * Run Electrophoresis test on digestion | ||
[[User:Trentay|Trentay]] 23:24, 7 July 2009 (UTC) | [[User:Trentay|Trentay]] 23:24, 7 July 2009 (UTC) | ||
| + | |||
| + | |||
| + | ''Craig and Clay'' | ||
| + | * Since the Eidenburg team was not able to detect the presence of limonene, after transformation, we will see if we are able to use more sensitive equipment to detect its presence. The synthesized sequence for the LIM1 gene will also be studied to see if a more optimized sequence exists for E. coli. It must be noted that the Eidenburg team also deleted a target sequence at the beginning of the LIM1 gene which may have proven to be detrimental to its expression. Furthermore, the pathway that the LIM1 gene competes with may be critical to cell survival. These possible reasons will be more carefully deconstructed. | ||
---- | ---- | ||
====Wetlab==== | ====Wetlab==== | ||
| - | '' | + | ''Maria and Afton'' |
* Digestion | * Digestion | ||
| - | |||
* Agarose Gel preparation/Electrophoresis test | * Agarose Gel preparation/Electrophoresis test | ||
| + | * Spectrometry of Miniprepped DNA | ||
| + | {| cellpadding="5" border="1" padding="5" | ||
| + | |+ | ||
| + | |- | ||
| + | ! Part No.!! A260 (nm) !! A280 (nm) !! A260/A280 !! Volume (µL) !! Conc.(µg/mL) !!Amount (µg) | ||
| + | |- | ||
| + | ! pSB3K3 (1) | ||
| + | | 0.011 || 0.010 || 1.10 || 35 || 11 || 0.385 | ||
| + | |- | ||
| + | ! pSB3K3 (2) | ||
| + | | 0.010 || 0.008 || 1.25 || 35 || 10 || 0.35 | ||
| + | |- | ||
| + | ! pSB3K3 (3) | ||
| + | | 0.010 || 0.008 || 1.25 || 30 || 10 || 0.30 | ||
| + | |} | ||
| + | * Since it is impossible to do a successful BioBrick assembly with this small amount of concentraiton we combined all 3 vials of pSB3K3 to create a vial containing 100uL of DNA, and the Measurements were retaken | ||
| + | {| cellpadding="5" border="1" padding="5" | ||
| + | |+ | ||
| + | |- | ||
| + | ! Part No.!! A260 (nm) !! A280 (nm) !! A260/A280 !! Volume (µL) !! Conc. (µg/mL) !!Amount (µg) | ||
| + | |- | ||
| + | ! pSB3K3 (All) | ||
| + | | 0.010 || 0.010 || 1.0 || 97 || 20 || 1.94 | ||
| + | |} | ||
[[User:Trentay|Trentay]] 21:29, 7 July 2009 (UTC) | [[User:Trentay|Trentay]] 21:29, 7 July 2009 (UTC) | ||
Latest revision as of 16:43, 13 August 2009
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Contents |
Tuesday 7 July 2009
Results
- Everyone is still waiting on electrotransformation cuvettes to proceed with projects
Maria and Afton
- Miniprep Results:
| Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg) |
|---|---|---|---|---|---|---|
| pSB3K3 (1) | 0.011 | 0.010 | 1.10 | 35 | 11 | 0.385 |
| pSB3K3 (2) | 0.010 | 0.008 | 1.25 | 35 | 10 | 0.350 |
| pSB3K3 (3) | 0.010 | 0.008 | 1.25 | 30 | 11 | 0.300 |
| pSB3K3 (All) | 0.010 | 0.010 | 1.00 | 97 | 20 | 1.940 |
- Individually the concentrations of pSB3K3 were too small for digestion, so all of the Miniprepped samples were combined and measurements were retaken.
- pSB3K3 backbone did not show up on the gel. Only the ladder is visible
Trentay 23:23, 7 July 2009 (UTC)
Tasks
- Continue research and writing paper drafts
Maria and Afton
- Take Spectrometer measurements to get DNA amounts of pSB3K3
- Digestion of pSB3K3
- Work on Excel record of stocks
- Run Electrophoresis test on digestion
Trentay 23:24, 7 July 2009 (UTC)
Craig and Clay
- Since the Eidenburg team was not able to detect the presence of limonene, after transformation, we will see if we are able to use more sensitive equipment to detect its presence. The synthesized sequence for the LIM1 gene will also be studied to see if a more optimized sequence exists for E. coli. It must be noted that the Eidenburg team also deleted a target sequence at the beginning of the LIM1 gene which may have proven to be detrimental to its expression. Furthermore, the pathway that the LIM1 gene competes with may be critical to cell survival. These possible reasons will be more carefully deconstructed.
Wetlab
Maria and Afton
- Digestion
- Agarose Gel preparation/Electrophoresis test
- Spectrometry of Miniprepped DNA
| Part No. | A260 (nm) | A280 (nm) | A260/A280 | Volume (µL) | Conc.(µg/mL) | Amount (µg) |
|---|---|---|---|---|---|---|
| pSB3K3 (1) | 0.011 | 0.010 | 1.10 | 35 | 11 | 0.385 |
| pSB3K3 (2) | 0.010 | 0.008 | 1.25 | 35 | 10 | 0.35 |
| pSB3K3 (3) | 0.010 | 0.008 | 1.25 | 30 | 10 | 0.30 |
- Since it is impossible to do a successful BioBrick assembly with this small amount of concentraiton we combined all 3 vials of pSB3K3 to create a vial containing 100uL of DNA, and the Measurements were retaken
| Part No. | A260 (nm) | A280 (nm) | A260/A280 | Volume (µL) | Conc. (µg/mL) | Amount (µg) |
|---|---|---|---|---|---|---|
| pSB3K3 (All) | 0.010 | 0.010 | 1.0 | 97 | 20 | 1.94 |
Trentay 21:29, 7 July 2009 (UTC)
