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Team:Hawaii/Notebook/2008-09- 6
From 2008.igem.org
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===OriT test=== | ===OriT test=== | ||
| - | <strong>Margaret</strong> | + | :<strong>Margaret</strong> |
:*The biobrick form of OriTr did not show up on the above mentioned gel, so I am: | :*The biobrick form of OriTr did not show up on the above mentioned gel, so I am: | ||
::*PCR amplifying it from the biobrick paper | ::*PCR amplifying it from the biobrick paper | ||
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===Culturing=== | ===Culturing=== | ||
| - | <strong>Margaret</strong> | + | :<strong>Margaret</strong> |
:*DH5-alpha (for making competent cells) | :*DH5-alpha (for making competent cells) | ||
:*rep, J33207, pSB3K3, and omega interposon | :*rep, J33207, pSB3K3, and omega interposon | ||
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= Discussion = | = Discussion = | ||
Latest revision as of 04:09, 10 September 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Construction of B_H_R Vector
- Margaret
- Yesterday's Restriction Digest
- Plac+B0030 and Plac+B0034: ethanol precipitation http://openwetware.org/wiki/Ethanol_precipitation_of_small_DNA_fragments
- pSB1A3, pSB3K3, J33207: de-phosphorylated using SAP
- Gel Quantification
- Some of these products did not appear on the gel, so other methods will be used to generate them.
- Plac+B0030 and Plac+B0034 did not show up on the gel, so the ligation is being performed tonight, pSB1A3 and pSB3K3 plasmid preps from 9/4 showed a good band.
- Ligation 1:1, followed by Restriction digest with E,S,& P (to cut up any polymers formed and to leave E & S overhang):
- Plac + B0030 (500ng)
- Plac + B0030 (100ng)
- Plac + B0034 (500ng)
- Plac + B0030 (100ng)
- pSB3K3 restriction digested with E, P
OriT test
- Margaret
- The biobrick form of OriTr did not show up on the above mentioned gel, so I am:
- PCR amplifying it from the biobrick paper
- Extracting DNA from the biobrick registry (1004, 10D) and transforming into DH5-alpha cells. The negative control is pSB1A3-dephosphorylated, the positive control is pSB1A3, from a plasmid prep.
Culturing
- Margaret
- DH5-alpha (for making competent cells)
- rep, J33207, pSB3K3, and omega interposon
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 
]
