Team:Michigan/Pili Expression
From 2010.igem.org
(Difference between revisions)
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!Week 9 | !Week 9 | ||
| [[Team:Michigan/Pili_Expression#8/22/2010|8/22/2010]] | | [[Team:Michigan/Pili_Expression#8/22/2010|8/22/2010]] | ||
- | | | + | | [[Team:Michigan/Pili_Expression#8/23/2010|8/23/2010]] |
| - | | - | ||
| - | | - | ||
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##use EB buffer as the blank | ##use EB buffer as the blank | ||
##ALWAYS clean twice when done using the machine | ##ALWAYS clean twice when done using the machine | ||
- | {| | + | |
+ | {| style="color:#1c2bf2;background-color:#fafa19;font-size:9pt;text-align:center" cellpadding="5" cellspacing="0" border="1" bordercolor="#fff" width="33%" | ||
|- | |- | ||
! | ! | ||
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Prepared culture of DH5a for electroporation tomorrow. | Prepared culture of DH5a for electroporation tomorrow. | ||
+ | |||
+ | ==8/23/2010== | ||
+ | ''Kevin, Marc'' | ||
+ | |||
+ | Growing competent cells for electroporation. | ||
+ | |||
+ | Colony started growing at 2:00. | ||
+ | |||
+ | Measured ODs | ||
+ | {| style="color:#1c2bf2;background-color:#fafa19;font-size:9pt;text-align:center" cellpadding="5" cellspacing="0" border="1" bordercolor="#fff" width="33%" | ||
+ | |- | ||
+ | ! Time | ||
+ | ! OD600 | ||
+ | |- | ||
+ | | 3:30 | ||
+ | | .114 | ||
+ | |- | ||
+ | | 5:30 | ||
+ | | .206 | ||
+ | |- | ||
+ | | 6:30 | ||
+ | | .441 | ||
+ | |} | ||
+ | |||
+ | Precipitate Ligation product w/butanol | ||
+ | |||
+ | Add to eppendorf tube: | ||
+ | #50 ul ultrapure H2O | ||
+ | #Ligation product | ||
+ | #500 ul butanol | ||
+ | Spin at 4C, 13000 rpm for 20 min | ||
+ | |||
+ | Made ampicillin plates | ||
+ | *Add 25 ul Amp100 to LB plates | ||
+ | |||
+ | 6 plates: | ||
+ | *Cells only | ||
+ | *Plasmid only | ||
+ | *2x pBAD+FimB undiluted | ||
+ | *2x pBAD+FimB 1:100 | ||
Revision as of 22:01, 23 August 2010