Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/03/12: Difference between revisions
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==Entry title== | ==Entry title== | ||
Stopped the sender cultures at noon. Spun in 2ml tubes for 3 minutes at 13.1g. Poured supernatants into 15ml conical tubes. Added 5ml LB to the round bottom tunes. Started shaking again at 1:30pm. | |||
Afternoon: | |||
Added 5ul of 50mM AHL to 5ml water, added 1ml of hexane to that, mixed vigorously, can also heat<br> | |||
Mess with ratio, can heat it also<br> | |||
Can centrifuge it down to help break it up<br> | |||
Move top layer directly to HPLC vial<br> | |||
Put vial in the right spot<br> | |||
Hit start<br> | |||
<br> | |||
GC<br> | |||
Open all 3 cylinders<br> | |||
Check pressures after turning the machines on<br> | |||
Turn on top Hewlett Packard thing<br> | |||
Turn on GC machine (in back on right side)<br> | |||
Open software<br> | |||
Open fiD air A detector<br> | |||
Close hydrogen<br> | |||
Push the FiD igniter (instructions on wall for this part)<br> | |||
Check that it's lit by looking down the hole using the wrench as a mirror and see if condensation forms on it<br> | |||
If any data ever looks funny, check that condensation still forms on the wrench<br> | |||
<br> | |||
Software | |||
<br> | |||
Sequence parameters<br> | |||
Signal 2 prefix<br> | |||
Make sure it goes to my directory<br> | |||
Method is AHLm<br> | |||
Flame ionization detector<br> | |||
Controls<br> | |||
5 step calibration curve<br> | |||
.5ml hexane<br> | |||
2ul<br> | |||
1.5ul<br> | |||
1ul<br> | |||
.5ul<br> | |||
.25ul<br> | |||
<br> | |||
Started at 340, 23 samples | |||
tonight: | tonight: | ||
Revision as of 16:21, 28 May 2013
Project name | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
Entry titleStopped the sender cultures at noon. Spun in 2ml tubes for 3 minutes at 13.1g. Poured supernatants into 15ml conical tubes. Added 5ml LB to the round bottom tunes. Started shaking again at 1:30pm.
Software
Sequence parameters Flame ionization detector
miniprepped E0420 (E0343 didn't grow) cut with X & P cut K092900 with S & P gel extracted ligated
3.8ul backbone 13ul insert behzad's roche buffer and ligase not doing backbone control
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