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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=" | + | {| cellpadding="5" border="1" padding="5" |
|+ | |+ | ||
|- | |- | ||
| - | ! Promoter!! Reporter !! | + | ! Promoter!! Reporter !! Plate !! Growth Observation |
|- | |- | ||
! J23101 | ! J23101 | ||
| - | | E0240 || Cm || no growth | + | | E0240 || Cm || no growth |
|- | |- | ||
! J23101 | ! J23101 | ||
| - | | J06702 || Cm || 21 colonies | + | | J06702 || Cm || 21 colonies |
|- | |- | ||
! J23102 | ! J23102 | ||
| - | | E0240 || Cm || 3 colonies | + | | E0240 || Cm || 3 colonies |
|- | |- | ||
! J23102 | ! J23102 | ||
| - | | J06702 || Cm || 8 colonies | + | | J06702 || Cm || 8 colonies |
|- | |- | ||
! J23103 | ! J23103 | ||
| - | | E0240 || Cm || 2 colonies | + | | E0240 || Cm || 2 colonies |
|- | |- | ||
! J23103 | ! J23103 | ||
| - | | J06702 || Cm || 23 colonies | + | | J06702 || Cm || 23 colonies |
|- | |- | ||
! J23104 | ! J23104 | ||
| - | | E0240 || Cm || 9 colonies | + | | E0240 || Cm || 9 colonies |
|- | |- | ||
! J23104 | ! J23104 | ||
| - | | J06702 || Cm || 12 colonies | + | | J06702 || Cm || 12 colonies |
|- | |- | ||
! J23105 | ! J23105 | ||
| - | | E0240 || Cm || no growth | + | | E0240 || Cm || no growth |
|- | |- | ||
! J23105 | ! J23105 | ||
| - | | J06702 || Cm || 14 colonies | + | | J06702 || Cm || 14 colonies |
|- | |- | ||
! J23107 | ! J23107 | ||
| - | | E0240 || Cm || 3 colonies | + | | E0240 || Cm || 3 colonies |
|- | |- | ||
! J23107 | ! J23107 | ||
| - | | J06702 || Cm || 3 colonies | + | | J06702 || Cm || 3 colonies |
|} | |} | ||
* 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) | ||
| - | + | ''Craig and Clay'' | |
* No growth was seen on either the kanamycin or chloramphenicol plates. | * 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=== | ===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 | ||
| Line 60: | Line 128: | ||
* Transformation of NEB 10 beta to make sure new stock is electrocompetent | * Transformation of NEB 10 beta to make sure new stock is electrocompetent | ||
[[User:Trentay|Trentay]] 20:41, 16 July 2009 (UTC) | [[User:Trentay|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. | ||
---- | ---- | ||
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)
