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Team:Hawaii/Notebook/2008-08-30
From 2008.igem.org
(Difference between revisions)
(→Construction of p+r and rereplacement of BB-pRL1383a MCS) |
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= Things we did today = | = Things we did today = | ||
== Wetlab work == | == Wetlab work == | ||
| - | ===Construction of p+r and | + | ===Construction of p+r and re-replacement of BB-pRL1383a MCS=== |
:<strong> Grace</strong> | :<strong> Grace</strong> | ||
[[Image:083008REdigests.jpg|right|thumb|400px|EtBr stained 2% (L) and 0.8% (R) agarose gels ran at 60V for 2 hours and 1 hour, respectively. Thirty microliters of RE digest reaction were loaded into each well.]] | [[Image:083008REdigests.jpg|right|thumb|400px|EtBr stained 2% (L) and 0.8% (R) agarose gels ran at 60V for 2 hours and 1 hour, respectively. Thirty microliters of RE digest reaction were loaded into each well.]] | ||
| Line 11: | Line 11: | ||
::* nir: ~800bp band (from GFP contaminant in plasmid prep, cut only once) and ~2kb band (??) | ::* nir: ~800bp band (from GFP contaminant in plasmid prep, cut only once) and ~2kb band (??) | ||
::* BB-pRL1383a: ran WAY too much product. Will redo RE digest with 20% of plasmid used. Since J33207 was only cut once, it's assumed BB-pRL1383a was cut only once as well. | ::* BB-pRL1383a: ran WAY too much product. Will redo RE digest with 20% of plasmid used. Since J33207 was only cut once, it's assumed BB-pRL1383a was cut only once as well. | ||
| - | :* Extracted | + | :* Extracted C0012 vector band from gel |
:* Ligated: | :* Ligated: | ||
::* plac + rbs (B0034) | ::* plac + rbs (B0034) | ||
Revision as of 01:32, 31 August 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Construction of p+r and re-replacement of BB-pRL1383a MCS
- Grace
- Ran RE digests from last night on agarose gel
- C0012: correct vector band ~2.1kb
- J33207: band at ~700bp (only one enzyme cut -- XbaI did not cut; PstI cuts because C0012 was cut by PstI) and ~850bp (uncut PCR product)
- nir: ~800bp band (from GFP contaminant in plasmid prep, cut only once) and ~2kb band (??)
- BB-pRL1383a: ran WAY too much product. Will redo RE digest with 20% of plasmid used. Since J33207 was only cut once, it's assumed BB-pRL1383a was cut only once as well.
- Extracted C0012 vector band from gel
- Ligated:
- plac + rbs (B0034)
- PCR of J33207
- RE digested:
- p+r with EcoRI, XbaI, SpeI, PstI
- J33207 and BB-pRL1383a with EcoRI and PstI
- Treated C0012 vector (pSB1A2) with SAP
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 
]
