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Minnesota/17 July 2008
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|6. '''Double digest of all ligations:''' from previous days. Do a double digest to cut the components linearly, which will then be able to run through a gel to check if correct DNA is present. Follow the table below: | |6. '''Double digest of all ligations:''' from previous days. Do a double digest to cut the components linearly, which will then be able to run through a gel to check if correct DNA is present. Follow the table below: | ||
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|7. '''Gel Electrophoresis:''' Performed on double digest reaction from above. Refer to picture below for results. | |7. '''Gel Electrophoresis:''' Performed on double digest reaction from above. Refer to picture below for results. | ||
|[[Image:7.17.08gel.jpg|thumb|left|600px|Gel from 07-17-2008]] | |[[Image:7.17.08gel.jpg|thumb|left|600px|Gel from 07-17-2008]] | ||
Revision as of 15:26, 18 July 2008
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| 1. Plasmid prep dual promoters: (1) Tet & P22mnt, (2) LacI & LAMBDAcI. | ||||||
| 2. Model: Figure out why have such a low GFP output when run model. | ||||||
| 3. Sequence L5, L6-L9 | ||||||
| 4. Re-transform RFP, TetR promoter, terminator, and MCherry because no cell growth on plates. Place in 2mL LB cultures, allow growth in incubator @37C for 2 hours. Plate samples on Ampicillin resistant plates. Allow growth O/N. | ||||||
| 5. Discuss problems with sequences | ||||||
| 6. Double digest of all ligations: from previous days. Do a double digest to cut the components linearly, which will then be able to run through a gel to check if correct DNA is present. Follow the table below: |
| Parts | 10x Buffer | BSA | H20 | DNA | RE 1 | RE 2 |
|---|---|---|---|---|---|---|
| L2a, LAMBDAcI:Term | 5.0uL | 0.5uL | 32.5uL | 10.0uL | EcoRI | Xba1 |
| L2b, LAMBDAcI:Term | 5.0uL | 0.5uL | 32.5uL | 10.0uL | EcoRI | Xba1 |
| L3i, Pro:LAMBDAcI | 5.0uL | 0.5uL | 32.5uL | 10.0uL | Pst1 | Spe1 |
| L3j, Pro:LAMBDAcI | 5.0uL | 0.5uL | 32.5uL | 10.0uL | Pst1 | Spe1 |
| L4b, LAMBDAcI:Term | 5.0uL | 0.5uL | 40.5uL | 2.0uL | EcoRI | Xba1 |
| L4c, LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1 |
| L4d, LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1 |
| L4e, LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1 |
| L4g, LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1 |
| L4h, LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1 |
| L4i, LAMBDAcI:Term | 5.0uL | 0.5uL | 37.5uL | 5.0uL | EcoRI | Xba1 |
| L4j, LAMBDAcI:Term | 5.0uL | 0.5uL | 32.5uL | 10.0uL | EcoRI | Xba1 |
| L5c, GFP:Term | 5.0uL | 0.5uL | 40.5uL | 2.0uL | EcorI | Xba1 |
| L6a, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1 |
| L6b, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 41.5uL | 1.0uL | EcoRI | Xba1 |
| L6e, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1 |
| L6d, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1 |
| L7a, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 38.5uL | 4.0uL | EcoRI | Xba1 |
| L7b, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1 |
| L7d, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 37.5uL | 5.0uL | EcoRI | Xba1 |
| L8a, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1 |
| L9a, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 41.5uL | 1.0uL | EcoRI | Xba1 |
| L9c, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 40.5uL | 2.0uL | EcoRI | Xba1 |
| L9d, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 41.5uL | 1.0uL | EcoRI | Xba1 |
