User:Daniel Goodman/Notebook/Cluzel/2010/02/22

Prep

 * 3% w/v agarose
 * 5 ml LB, 150 mg agarose
 * heat in 80 deg. bath for 15 min
 * 108 ul per mold


 * 6 agarose block

Method

 * 1) Mold agarose into blocks
 * 2) Not using silicon molds, only testing bacterial dispersion on agarose/glass slide.
 * 3) waiting 20 min for agarose to dry in PDMS molds, at room temp; in future, will try different drying temps/dessication?
 * 4) cover agarose in mold with cover slip
 * 5) Place all 6 blocks on rectangular glass slides


 * 1) Grew up wild-type MG1655 E. coli to exponential phase, 0.1 ul of solution (measure OD)
 * 2) spot two 0.2 ul drops, one on each end of each agarose block/corresp. slide position, wait requisite time for each

Varying:
 * wait time
 * 1 min, 5 min, 10 min
 * deposition surface
 * agarose: dessicate agarose for 1, 5, 10 min, then add 0.2 ul of cells, then cover slip
 * glass coverslip, transfer to agarose: dry cells for 1, 5, 10 min on cover slip, then xfer

Measuring:
 * spot dispersion diameter

Data
'''Fail: Two agarose blocks per slide causes the objective to move away, making focusing impossible. Redo next time with one agarose block per slide.'''


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