User:Steven J. Koch/Notebook/Kochlab/2009/06/15/TRITC (Rhodamine) and regular tubulin

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See also Linh's notes: User:Linh N Le/Notebook/2009/06/15

  1. Make GPEM
  2. Make GPEM + Cushion (1/2 cushion?)
  3. Resuspend one aliquot of TRITC tubulin with 4 ul of GPEM+Cushion. Make 3 aliquots; flash-freeze
    • One freeze-dried aliquot is $25-ish, so each 1 ul aliquot is about $6.
  4. Resuspend one aliquot of unlabeled tubulin with 200 ul of GPEM + cushion. Make ___ number of aliquots.
    • One freeze-dried aliquot is $95. If we made 5 ul aliquots (40 total), that would be about $2.5 / new aliquot.

Contents

Making GPEM

Steve Koch 16:38, 15 June 2009 (EDT): Linh is making GPEM. He is making 300 microliters total (294 ul PEM + 6 ul GTP)

Making GPEM + Cushion

Steve Koch 16:46, 15 June 2009 (EDT): When resuspending labeled tubulin aliquot, need to mix up GPEM + cushion, because it's easier than trying to add a small amount of cushion after resuspension.

  • 9 microliters GPEM
  • 1 microliter Cushion (from cytoskeleton)

Resuspending rhodamine tubulin aliquot

Resuspending unlabeled tubulin aliquot

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