Virginia Commonwealth/16 July 2009
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==Thursday 16 July 2009== | ==Thursday 16 July 2009== | ||
===Results=== | ===Results=== | ||
+ | ''Maria and Afton'' | ||
* Results of Retransformation with pSB4C5 backbone in NEB 10 beta two days after transformation | * Results of Retransformation with pSB4C5 backbone in NEB 10 beta two days after transformation | ||
{| cellpadding="5" border="1" padding="5" | {| cellpadding="5" border="1" padding="5" | ||
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* No growth was seen in 5mL culture taken from NEB 10 beta cell freezer stock | * No growth was seen in 5mL culture taken from NEB 10 beta cell freezer stock | ||
**Vial may have thawed out too much after cells were taken out for the electrocompetency procedure | **Vial may have thawed out too much after cells were taken out for the electrocompetency procedure | ||
+ | |||
+ | * Spectrophotometry measurements after Miniprep | ||
+ | {| cellpadding="5" border="1" padding="5" | ||
+ | |+ | ||
+ | |- | ||
+ | ! Part No.!! A260 (nm) !! A280 (nm) !! A260/A280 !! Vol. (µL) !! Conc. (µg/mL) !! Amt (µg) | ||
+ | |- | ||
+ | | J23110 || 0.050 || 0.038 || 1.32 || 48 || 75 || 3.60 | ||
+ | |- | ||
+ | | J23113 || 0.041 || 0.030 || 1.37 || 44 || 21 || 0.92 | ||
+ | |- | ||
+ | | J23116 || 0.029 || 0.023 || 1.26 || 44 || 43 || 1.91 | ||
+ | |- | ||
+ | | R0011 || 0.022 || 0.020 || 1.10 || 41 || 33 || 1.35 | ||
+ | |- | ||
+ | | R0040 || 0.024 || 0.021 || 1.14 || 42 || 36 || 1.51 | ||
+ | |- | ||
+ | | E0240 || 0.056 || 0.036 || 1.56 || 109 || 84 || 9.16 | ||
+ | |- | ||
+ | | pSB4C5 || 0.116 || 0.066 || 1.76 || 174 || 22 || 3.83 | ||
+ | |- | ||
+ | | J06702 || 0.042 || 0.024 || 1.75 || 63 || 130 || 8.19 | ||
+ | |} | ||
+ | |||
+ | * Gel Electrophoresis Results | ||
+ | [[Image:2009-07-16 digest07 110,113,116,0011,0040,240.jpg|350px|thumb|none| 100bp Ladder, Digestions on the Gel: J23110, J23113, J23116, R0011, R0040, E0240]] | ||
+ | |||
[[User:Trentay|Trentay]] 14:31, 16 July 2009 (UTC) | [[User:Trentay|Trentay]] 14:31, 16 July 2009 (UTC) | ||
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===Tasks=== | ===Tasks=== | ||
''Maria and Afton'' | ''Maria and Afton'' | ||
+ | * Spectrophotometry | ||
* Digest: E0240,J06702, pSB4C5, 110, 113, 116, 0011, 0040 | * Digest: E0240,J06702, pSB4C5, 110, 113, 116, 0011, 0040 | ||
* Ligate promoters with backbone pSB4C5 and reporters E0240, J06702 | * Ligate promoters with backbone pSB4C5 and reporters E0240, J06702 |
Latest revision as of 18:17, 13 August 2009
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Contents |
Thursday 16 July 2009
Results
Maria and Afton
- Results of Retransformation with pSB4C5 backbone in NEB 10 beta two days after transformation
Promoter | Reporter | Plate | Growth Observation |
---|---|---|---|
J23101 | E0240 | Cm | no growth |
J23101 | J06702 | Cm | 21 colonies |
J23102 | E0240 | Cm | 3 colonies |
J23102 | J06702 | Cm | 8 colonies |
J23103 | E0240 | Cm | 2 colonies |
J23103 | J06702 | Cm | 23 colonies |
J23104 | E0240 | Cm | 9 colonies |
J23104 | J06702 | Cm | 12 colonies |
J23105 | E0240 | Cm | no growth |
J23105 | J06702 | Cm | 14 colonies |
J23107 | E0240 | Cm | 3 colonies |
J23107 | J06702 | Cm | 3 colonies |
- No growth was seen in 5mL culture taken from NEB 10 beta cell freezer stock
- Vial may have thawed out too much after cells were taken out for the electrocompetency procedure
- Spectrophotometry measurements after Miniprep
Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg) |
---|---|---|---|---|---|---|
J23110 | 0.050 | 0.038 | 1.32 | 48 | 75 | 3.60 |
J23113 | 0.041 | 0.030 | 1.37 | 44 | 21 | 0.92 |
J23116 | 0.029 | 0.023 | 1.26 | 44 | 43 | 1.91 |
R0011 | 0.022 | 0.020 | 1.10 | 41 | 33 | 1.35 |
R0040 | 0.024 | 0.021 | 1.14 | 42 | 36 | 1.51 |
E0240 | 0.056 | 0.036 | 1.56 | 109 | 84 | 9.16 |
pSB4C5 | 0.116 | 0.066 | 1.76 | 174 | 22 | 3.83 |
J06702 | 0.042 | 0.024 | 1.75 | 63 | 130 | 8.19 |
- Gel Electrophoresis Results
Trentay 14:31, 16 July 2009 (UTC)
Craig and Clay
- No growth was seen on either the kanamycin or chloramphenicol plates.
- The sequence for the LIM1 gene was verified to be optimized for E. coli.
Tasks
Maria and Afton
- Spectrophotometry
- Digest: E0240,J06702, pSB4C5, 110, 113, 116, 0011, 0040
- Ligate promoters with backbone pSB4C5 and reporters E0240, J06702
- Run an electrophoresis test on promoters and reporter Eo240
- Pick colonies of promoters from transformation 100-107 (excluding 106)
- Transformation of NEB 10 beta to make sure new stock is electrocompetent
Trentay 20:41, 16 July 2009 (UTC)
Craig and Clay
- The registry will be contacted to verify the correct resistence for the limonene synthesis related parts. New DNA will also be requested.
Wetlab
Maria and Afton
- Digest: E0240,J06702, pSB4C5, 110, 113, 116, 0011, 0040
- Ligate promoters with backbone pSB4C5 and reporters E0240, J06702
- Run an electrophoresis test on promoters and reporter Eo240
Trentay 20:41, 16 July 2009 (UTC)