Virginia Commonwealth/14 July 2009
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===Results=== | ===Results=== | ||
''Maria and Afton'' | ''Maria and Afton'' | ||
- | * (+) | + | * All of the promoters showed up on the gel electrophoresis in a mostly straight line. |
- | + | * Transformation Results | |
- | + | {| cellpadding="5" border="1" padding="5" | |
- | + | |+ | |
- | + | |- | |
- | + | ! Name !! Plate !! Growth Observation | |
- | + | |- | |
- | + | | Control (+) NEB10β || LB || lawn | |
+ | |- | ||
+ | | Control (+) NEB10β shocked || LB || lawn | ||
+ | |- | ||
+ | | Control (-) NEB10β || Cm || no growth | ||
+ | |- | ||
+ | | J23106, J06702, pSB4C5 ligation || Cm || ~30 small colonies | ||
+ | |} | ||
+ | |||
+ | * Miniprep Results: | ||
+ | {| cellpadding="5" border="1" padding="5" | ||
+ | |+ | ||
+ | |- | ||
+ | ! Part No.!! A260 (nm) !! A280 (nm) !! A260/A280 !! Vol. (µL) !! Conc. (µg/mL) !! Amt (µg) | ||
+ | |- | ||
+ | | J23101 || 0.050 || 0.039 || 1.20 || 47 || 75 || 4.70 | ||
+ | |- | ||
+ | | J23102 || 0.030 || 0.025 || 1.20 || 58 || 45 || 3.48 | ||
+ | |- | ||
+ | | J23103 || 0.044 || 0.032 || 1.38 || 56 || 66 || 4.93 | ||
+ | |- | ||
+ | | J23104 || 0.033 || 0.028 || 1.18 || 54 || 49 || 3.56 | ||
+ | |- | ||
+ | | J23105 || 0.037 || 0.029 || 1.28 || 32 || 56 || 1.78 | ||
+ | |- | ||
+ | | J23107 || 0.054 || 0.040 || 1.35 || 47 || 81 || 3.81 | ||
+ | |} | ||
[[User:Trentay|Trentay]] 13:44, 14 July 2009 (UTC) | [[User:Trentay|Trentay]] 13:44, 14 July 2009 (UTC) | ||
+ | |||
+ | * Gel Electrophoresis Results | ||
+ | [[Image:2009-07-14 digest06 100-105,107.jpg|350px|thumb|none|from left to right: ladder, 100-105,107]] | ||
+ | |||
''Kevin and Adam'' | ''Kevin and Adam'' | ||
*Miniprep results: | *Miniprep results: | ||
+ | {| cellpadding="5" border="1" padding="5" | ||
+ | |+ | ||
+ | |- | ||
+ | ! Part No.!! A260 (nm) !! A280 (nm) !! A260/A280 !! Vol. (µL) !! Conc. (µg/mL) !! Amt (µg) | ||
+ | |- | ||
+ | |E0240||.033||.019||1.737||124||66||8.184 | ||
+ | |- | ||
+ | |pSB4C5||.054||.036||1.5||123.5||108||13.34 | ||
+ | |} | ||
[[User:Bussingkm|Bussingkm]] 15:21, 14 July 2009 (UTC) | [[User:Bussingkm|Bussingkm]] 15:21, 14 July 2009 (UTC) | ||
+ | |||
+ | ''Craig and Clay'' | ||
+ | * The ampicillin plates left overnight again displayed no growth. The electrotransformed parts will be re-plated onto ampicillin-selective plates as well as non-selective plates. | ||
---- | ---- | ||
Line 59: | Line 101: | ||
*Possible Ligation of promoters with parts J06702, pSB4C5 | *Possible Ligation of promoters with parts J06702, pSB4C5 | ||
[[User:Trentay|Trentay]] 15:13, 14 July 2009 (UTC) | [[User:Trentay|Trentay]] 15:13, 14 July 2009 (UTC) | ||
+ | |||
''Kevin and Adam'' | ''Kevin and Adam'' | ||
+ | *perform complete digestion and ligation | ||
*construct biobrick device (pSB4C5, BBa_E0240, BBa_J23100), and (pSB4C5, BBa_E0240, BBa_J23106) | *construct biobrick device (pSB4C5, BBa_E0240, BBa_J23100), and (pSB4C5, BBa_E0240, BBa_J23106) | ||
[[User:Bussingkm|Bussingkm]] 15:20, 14 July 2009 (UTC) | [[User:Bussingkm|Bussingkm]] 15:20, 14 July 2009 (UTC) | ||
- | --- | + | |
+ | ''Craig and Clay'' | ||
+ | * The electrotransformed parts will be re-plated on ampicillin plates as well as on non-selective plates. Growth on non-selective plates will prove the existence of viable cells. | ||
---- | ---- | ||
====Wetlab==== | ====Wetlab==== | ||
''Maria and Afton'' | ''Maria and Afton'' | ||
- | * Spectophotometry of promoters: 101, 102, 103, 104 | + | * Spectophotometry of promoters: 101, 102, 103, 104, 105, 107 |
- | * Digestion of promoters | + | * Digestion/Ligation of promoters |
- | * Electrophoresis of | + | * Electrophoresis of promoter digestions: 101, 102, 103, 104, 105, 107, (100) |
- | [[User:Trentay|Trentay]] | + | * Electrotransformation of ligated parts |
+ | ** two Cm plate had an unidentified growth in the refrigerator. | ||
+ | ** however, negative controls from the same plate stock show antibiotic is working | ||
+ | * Picked colonies for overnight culture of J23106 w/ J06702 reporter | ||
+ | * Made 3 overnight 5mL cultures of NEB 10 beta cell stock to prepare electrocompetent cells | ||
+ | [[User:Trentay|Trentay]] 21:01, 14 July 2009 (UTC) | ||
+ | |||
''Kevin and Adam'' | ''Kevin and Adam'' | ||
*pSB4C5, BBa_E0240, and BBa_J06702 were frozen in box C-11 of the glycerol stocks | *pSB4C5, BBa_E0240, and BBa_J06702 were frozen in box C-11 of the glycerol stocks | ||
[[User:Bussingkm|Bussingkm]] 15:22, 14 July 2009 (UTC) | [[User:Bussingkm|Bussingkm]] 15:22, 14 July 2009 (UTC) | ||
+ | |||
+ | ''Craig and Clay'' | ||
+ | * The electrotransformed parts were again plated on ampicillin-selective plates. They were also plated on non-selective media. One hundred microliters was used for each plate rather than the standard 35 microliters. |
Latest revision as of 20:45, 17 October 2009
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Contents |
Tuesday 14 July 2009
Results
Maria and Afton
- All of the promoters showed up on the gel electrophoresis in a mostly straight line.
- Transformation Results
Name | Plate | Growth Observation |
---|---|---|
Control (+) NEB10β | LB | lawn |
Control (+) NEB10β shocked | LB | lawn |
Control (-) NEB10β | Cm | no growth |
J23106, J06702, pSB4C5 ligation | Cm | ~30 small colonies |
- Miniprep Results:
Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg) |
---|---|---|---|---|---|---|
J23101 | 0.050 | 0.039 | 1.20 | 47 | 75 | 4.70 |
J23102 | 0.030 | 0.025 | 1.20 | 58 | 45 | 3.48 |
J23103 | 0.044 | 0.032 | 1.38 | 56 | 66 | 4.93 |
J23104 | 0.033 | 0.028 | 1.18 | 54 | 49 | 3.56 |
J23105 | 0.037 | 0.029 | 1.28 | 32 | 56 | 1.78 |
J23107 | 0.054 | 0.040 | 1.35 | 47 | 81 | 3.81 |
Trentay 13:44, 14 July 2009 (UTC)
- Gel Electrophoresis Results
Kevin and Adam
- Miniprep results:
Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg) |
---|---|---|---|---|---|---|
E0240 | .033 | .019 | 1.737 | 124 | 66 | 8.184 |
pSB4C5 | .054 | .036 | 1.5 | 123.5 | 108 | 13.34 |
Bussingkm 15:21, 14 July 2009 (UTC)
Craig and Clay
- The ampicillin plates left overnight again displayed no growth. The electrotransformed parts will be re-plated onto ampicillin-selective plates as well as non-selective plates.
Tasks
Maria and Afton
- Digest promoters: 101, 102, 103, 104
- Possible Ligation of promoters with parts J06702, pSB4C5
Trentay 15:13, 14 July 2009 (UTC)
Kevin and Adam
- perform complete digestion and ligation
- construct biobrick device (pSB4C5, BBa_E0240, BBa_J23100), and (pSB4C5, BBa_E0240, BBa_J23106)
Bussingkm 15:20, 14 July 2009 (UTC)
Craig and Clay
- The electrotransformed parts will be re-plated on ampicillin plates as well as on non-selective plates. Growth on non-selective plates will prove the existence of viable cells.
Wetlab
Maria and Afton
- Spectophotometry of promoters: 101, 102, 103, 104, 105, 107
- Digestion/Ligation of promoters
- Electrophoresis of promoter digestions: 101, 102, 103, 104, 105, 107, (100)
- Electrotransformation of ligated parts
- two Cm plate had an unidentified growth in the refrigerator.
- however, negative controls from the same plate stock show antibiotic is working
- Picked colonies for overnight culture of J23106 w/ J06702 reporter
- Made 3 overnight 5mL cultures of NEB 10 beta cell stock to prepare electrocompetent cells
Trentay 21:01, 14 July 2009 (UTC)
Kevin and Adam
- pSB4C5, BBa_E0240, and BBa_J06702 were frozen in box C-11 of the glycerol stocks
Bussingkm 15:22, 14 July 2009 (UTC)
Craig and Clay
- The electrotransformed parts were again plated on ampicillin-selective plates. They were also plated on non-selective media. One hundred microliters was used for each plate rather than the standard 35 microliters.