User:Daniel Goodman/Notebook/Cluzel/2010/03/05: Difference between revisions
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== | ==Grow cells== | ||
* | * Took a colony from fridge plate of MG1655 w/ GFP, put in 4 mL LB with 4 uL amp in 37°C shaker. (~10:30 AM) | ||
==Made agarose== | |||
* Made 5 mL of 5% agarose, and 5 mL of 3% agarose. (~10:40 AM) | |||
==Make chip impression in solidified gel== | |||
* Try quick experiment based on results from Wednesday; can we make features on a gel by pressing down? | |||
''Results: no features found with significant 'push' on 3% gel.'' | |||
==Try transferring again (w/ features)== | |||
*Scope access at 4PM | |||
*Start gel pouring at 2:30 - Poured 3% at 2:30, poured 5% at 2:50 | |||
===Notes=== | |||
*Not enough gel poured in mold, features did not form (but still continued w/ experiment) | |||
** In future, try to use more than 108, maybe 110. Removing bubbles (which happens often) often reduces height of agarose blob in mold and ruins features. | |||
*3% - couldn't find any cells, this could be because surface was concave so cells did not touch agarose | |||
*5% - cells found. Large (several screens), toward bottom of mold, so hard to tell extent - ''but clear boundaries were apparent!'' | |||
**Images saved in dbg, under 3-5-(5pct-gfp/phase)* | |||
Placement: | |||
#Used blade on handle to pry gel from out of mold so mold sat on surface of blade | |||
#Then used tweezers to grab long sides of mold from off of cell-coverslip, moved to slide to be imaged | |||
Next time: | |||
*Try spotting smaller amount; maybe using a needle, just a pipette tip (without taking up liquid) or something similar (nanodrop?). | |||
*Make journals for metamorph so multiple screen can be taken automatically | |||
*Figure out how stage increments in metamorph maps to nm/μm | |||
** Saved some slide positions for the blob in my blue notebook, maybe plot them clumsily later when we figure out slide position mapping | |||
==While waiting...== | |||
*Play with a few featureless gels, practice flipping & transferring | |||
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Latest revision as of 15:46, 5 March 2010
Cluzel Lab Notebook Daniel Goodman |
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Grow cells
Made agarose
Make chip impression in solidified gel
Results: no features found with significant 'push' on 3% gel. Try transferring again (w/ features)
Notes
Placement:
Next time:
While waiting...
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