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Team:Hawaii/Notebook/2008-09-24
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::* PCR machine froze while ExoSAPing. Unable to retrieve samples from machine. Jamie says she will figure it out. | ::* PCR machine froze while ExoSAPing. Unable to retrieve samples from machine. Jamie says she will figure it out. | ||
| - | ===Making Emergency BB Vector Insert (contaminant troubleshoot)=== | + | ===Making Emergency BB Vector Insert (help contaminant troubleshoot)=== |
[[Image:20080924-emergency_vector_pRLBB_vector_insert_PCR_redo.annotated.jpg|thumb|right|making x6 Parallel MCS Replacement fragmenets via PCR Redo!!! with clean uncontaminated primers [[Media:20080924-emergency_vector_pRLBB_vector_insert_PCR_redo.jpg|download unannotated]]]] | [[Image:20080924-emergency_vector_pRLBB_vector_insert_PCR_redo.annotated.jpg|thumb|right|making x6 Parallel MCS Replacement fragmenets via PCR Redo!!! with clean uncontaminated primers [[Media:20080924-emergency_vector_pRLBB_vector_insert_PCR_redo.jpg|download unannotated]]]] | ||
:<strong>Norman</strong> | :<strong>Norman</strong> | ||
Latest revision as of 09:31, 26 September 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Verified plasmid preps
- Grace
- PCR of plasmid preps from yesterday to verify inserts
- Ran PCR products on gel
- nrg #1 and nrsg #2 returned blanks. Bad plasmid prep?
- BB-pRL1383a and colony PCR of BB-pRL1383a E. coli both blank. Yikes!
- Quantified BB-pRL1383a and pSB1A3 plasmid preps
- Blanks. Really low conc.? Will nanodrop to check.
Construction of secretion device (cont.)
- Grace
- Overnight RE digest:
- nir, nrsg, and nrg PCR products with EcoRI and SpeI
- rgt PCR products with XbaI and PstI
- J33207 PCR product and pRL1383a plasmid with HindIII and BamHI
Prep for sequencing
- Grace
- nrg #4, nrg #7, nrsg #8, rgt #1, rgt #2
- plac+rbs
- PCR machine froze while ExoSAPing. Unable to retrieve samples from machine. Jamie says she will figure it out.
Making Emergency BB Vector Insert (help contaminant troubleshoot)
- Norman
- Cross posted in 2008-09-23 for comparison with contaminated PCR gel run.
Discussion
Quote of the Day
History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson
[http://manoa.hawaii.edu/ 
][http://manoa.hawaii.edu/ovcrge/ 
][http://www.ctahr.hawaii.edu 
]
