IGEM:IMPERIAL/2006/LabCalendar/2006-8-30: Difference between revisions
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*Part J37024 glass milk purified and electroporated. | *Part J37024 glass milk purified and electroporated. | ||
==Minipreps== | ==Minipreps of J37025, J37022, J37018== | ||
*See [[image:Miniprep_30-08.pdf|Miniprep_30-08.pdf|here]] for photo of gel | *See [[image:Miniprep_30-08.pdf|Miniprep_30-08.pdf|here]] for photo of gel | ||
*Comment: The part outlined in the photo as J37020 is actually J37022 !! | |||
==LoxP PCR== | ==LoxP PCR== | ||
| Line 51: | Line 52: | ||
*Ran along side the maxiprepped DNA | *Ran along side the maxiprepped DNA | ||
*Loaded 10{{uL}} of each PCR reaction into the gel (double the amount of the previous attempts) | *Loaded 10{{uL}} of each PCR reaction into the gel (double the amount of the previous attempts) | ||
==Freezing Cells== | |||
*Froze S01656 and J37020 (the right ones this time - old ones have been thrown out) | |||
==Culturing== | |||
*J37024 (from both plates for miniprep) | |||
*T9002/J37016/J37020/J37015/J37015RS/S01656 (for testing) | |||
Latest revision as of 09:14, 7 September 2006
TO DO: *Make Competent Cells *Electroporate J37015, J37024 (from ligations) after having made competent cells *Miniprep J37024, J37022 and J37025 (to check whether ligations were successful) *Religation of J37018 ? *Freeze 4G (S01656), J37020 (&J37018?) *Testing (T9002, J37016 & J37020) *PCR
Competent Cells
- New competent cells were made - we have a new stock in the -80[[:Category:{{{1}}}|{{{1}}}]] freezer again
Measuring the growth rate of the prey cells
- JohnChattaway 04:50, 30 August 2006 (EDT)
- We need to do this to work out the growth rate of the prey to give us an idea of the amounts of media we need for the chemostat.IGEM:IMPERIAL/2006/The Talk Page
- Cultured fresh cells at 9.30
- At 11.30 will re-culture to OD of 0.1 then test OD every 30mins untill OD 0.9 is reached
Measuring AHL production overnight
- JohnChattaway 04:50, 30 August 2006 (EDT)
- I will repeat tom's experiment with the frozen Prey Cells
Testing T9002, J37016 and J37020
- Took 16mL cultures out of shaker after 2h at 11:20
- OD measurements:
- T9002 0.186
- J37016 0.218
- J37020 0.160
- Dilute down to OD 0.1 into 25mL
- Put 5mL tubes with AHL in shaker at 12:00 - to be taken out at 16:00
- Tubes were taken out at 16:25
- Fluorescence reading of the 3 plates (17:50 - 19: 10)
Ligations
- Part J37024 glass milk purified and electroporated.
Minipreps of J37025, J37022, J37018
- See File:Miniprep 30-08.pdf for photo of gel
- Comment: The part outlined in the photo as J37020 is actually J37022 !!
LoxP PCR
- With any luck this should be the final attempt
- Ran along side the maxiprepped DNA
- Loaded 10μL of each PCR reaction into the gel (double the amount of the previous attempts)
Freezing Cells
- Froze S01656 and J37020 (the right ones this time - old ones have been thrown out)
Culturing
- J37024 (from both plates for miniprep)
- T9002/J37016/J37020/J37015/J37015RS/S01656 (for testing)
