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Virginia Commonwealth/30 July 2009
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''Maria and Afton'' | ''Maria and Afton'' | ||
* The overnight culture grew, however, no RFP was expressed (I1352) | * The overnight culture grew, however, no RFP was expressed (I1352) | ||
| + | * Spectrophotometry measurements | ||
| + | {| cellpadding="5" border="1" padding="5" | ||
| + | |+ | ||
| + | |- | ||
| + | ! Part No.!! A260 (nm) !! A280 (nm) !! A260/A280 !! Vol. (µL) !! Conc. (µg/mL) !! Amt (µg) | ||
| + | |- | ||
| + | | pSB4C5 || 0.125 || 0.076 || 1.65 || 27 || 250 || 6.75 | ||
| + | |- | ||
| + | | I1352 || 0.017 || 0.011 || 1.55 || 96 || 34 || 3.26 | ||
| + | |} | ||
| + | |||
| + | * Gel Electrophoresis Results | ||
| + | [[Image:2009-07-30 DigestI1352andpSB4C5.jpg|350px|thumb|none|from left to right: pSB4C5, I1352, 100bp Ladder, Bioline Hyperladder]] | ||
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[[User:Trentay|Trentay]] 14:26, 31 July 2009 (UTC) | [[User:Trentay|Trentay]] 14:26, 31 July 2009 (UTC) | ||
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===Tasks=== | ===Tasks=== | ||
''Maria and Afton'' | ''Maria and Afton'' | ||
Latest revision as of 18:16, 13 August 2009
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Contents |
Thursday 30 July 2009
Results
Maria and Afton
- The overnight culture grew, however, no RFP was expressed (I1352)
- Spectrophotometry measurements
| Part No. | A260 (nm) | A280 (nm) | A260/A280 | Vol. (µL) | Conc. (µg/mL) | Amt (µg) |
|---|---|---|---|---|---|---|
| pSB4C5 | 0.125 | 0.076 | 1.65 | 27 | 250 | 6.75 |
| I1352 | 0.017 | 0.011 | 1.55 | 96 | 34 | 3.26 |
- Gel Electrophoresis Results
Trentay 14:26, 31 July 2009 (UTC)
Tasks
Maria and Afton
- Make Cryogenic Stocks of part I1352 (2 vials)
- Make another overnight culture from the previous culture to see if it will express RFP
- Miniprep I1352
- Take spectrophotometry measurements of I1352 and pSB4C5
- Digest and Ligate I1352 and pSB4C5
- Run a Gel Electrophoresis on pSB4C5 and I1352 using the Hyper Ladder and the 100bp ladder
- Transform parts into NEB 10 beta
Trentay 14:26, 31 July 2009 (UTC)
Wetlab
Maria and Afton
- Cryogenic stocks were made
- Overnight culture of I1352 was grown up in 5mL of LB+AMP
- 2 samples of 1.5mL of I1352 was Miniprepped
- Spectophotometry measurements were taken
- The two samples were combined into one tube before measurements were taken
- pSB4C5 that was miniprepped on July 27, 2009
- Digestion was done
- Only the restriction enzymes EcoRI-HF and Pstl were used to cut the separate the backbone from the promter and RET
- Ligation was done
- for the Ligation of I1352 with pSB4C5, 3 microliters was added of each digestion to the Ligation sample
- for the Ligation of I1352 with itself, 6 microliters of the Digestion was addes to the Ligation sample
- Gel Electrophoresis was run at 120v for 45 min and DNA bands appeared for both samples
- Transformation was done on the two ligated samples
Trentay 15:10, 31 July 2009 (UTC)
