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Team:Stanford/Notebook/Lab Work/Week 5
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set up the following ligations from the gel extractions done by me, Karina, and Francisco on Friday, 7/14/10) | set up the following ligations from the gel extractions done by me, Karina, and Francisco on Friday, 7/14/10) | ||
*Francisco ran and imaged diagnostic gel on Friday | *Francisco ran and imaged diagnostic gel on Friday | ||
Revision as of 23:15, 20 July 2010
| Home | Project | Applications | Modeling | Parts | Team | Notebook |
Spring: Brainstorming | Spring Meetings
Summer: Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 | Week 7 | Week 8 | Week 9 | Week 10 | Summaries
|
7/19 Monday
Laura's Notebook
set up the following ligations from the gel extractions done by me, Karina, and Francisco on Friday, 7/14/10)
- Francisco ran and imaged diagnostic gel on Friday
| Ligation Recipe | |
|---|---|
| dH2O | none |
| vector (with terminators attached) | 5.0 uL |
| insert (RFP or GFP) | 12.0 uL |
| 10X buffer | 2.0 uL |
| T4 ligase | 1.0 uL |
- normally run for 10 minutes at room temperature, overnight this time (started at 11:30 am)
Nanodrop data (deemed unreliable based on 260/230, possibly due do residual EtBr contamination)
| part | 260/280 | 260/230 | ng/uL |
| vector/terminator | 1.77 | 0.03 | 19.1 |
| RFP | 1.86 | 0.02 | 16.2 |
| GFP | 1.83 | 0.02 | 14.8 |
7/20 Tuesday
Laura's Lab Notebook
helped Alex with preparation of competent cells
- see protocol: [http://openwetware.org/wiki/Stanford/BIOE44:Module_1:Day3 Preparing Electrocompetent Cells]
ran diagnostic gel for PCR-assembled DNA (Karina set up PCR rxns yesterday)
- order on gel:
- 100 bp ladder
- RSID1/RBS
- RSID2/RBS
