User:Linh N Le/Notebook/2009/06/23

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To Do
12-6
 * Koch/Atlas group meeting at 3
 * Try out new buffer we made yesterday?
 * Try Kinesin?
 * Clean up lab

BRB80

 * Since there are so many ways to make this buffer, Koch wanted me to try to find different famous people's recipes for the buffer
 * People to look for, Block and Vale
 * What we use for Cytoskeleton is called PEM and while PEM and BRB80 can be the same, they do not have to be
 * I did searches on both PEM and BRB80 and the following recipes are also reported by what they are called
 * Special Note: All PDF's linked here are from the personal website of the authors and are free access

Recipes

 * Hancock (Used by us as well)
 * 80 mM PIPES pH 6.85 (we use ph6.9)
 * 1 mM MgCl2
 * 1 mM EGTA
 * T.Salmon Labs
 * PM Buffer
 * 100 mM PIPES, pH 6.9
 * 2 mM EGTA
 * 1 mM Mg2SO4
 * Goldman Lab (Northwestern University)
 * 100 mM Pipes,pH 6.9
 * 1 mM MgCl2
 * 1 mM EGTA
 * Cold Spring Harbor Labs
 * PEM Buffer
 * 100 mM PIPES (pH 6.95)
 * 2 mM EGTA
 * 1 mM MgSO4
 * Cytoskeleton
 * PEM (General Tubulin Buffer)
 * 80 mM Na-PIPES pH 6.9
 * 1 mM MgCl2
 * 1 mM EGTA
 * Mitchison Lab (Harvard)
 * Brinkley Buffer 1980 (BRB80)
 * 80mM PIPES pH 6.8
 * 1mM MgCl2
 * 1mM EGTA
 * "Cytoskeleton Buffer"
 * 10mM MES pH 6.1
 * 138mM KCl
 * 3mM MgCl
 * 2mM EGTA
 * Notes: BRB80 is good for microtubules and 'cytoskeleton buffer' is good for both actin filaments and microtubules
 * Sigma Aldrich
 * PEM Buffer:
 * 0.1M PIPES (Product No. P8203)
 * 5 mM EGTA (Product No. E4378)
 * 2mM MgCl2 · 6H2O (Product No. M0250)
 * Bring to pH 6.8, using NaOH solution
 * Bayer College of Medicine
 * PEM Buffer:
 * 80mMPotassium PIPES, pH 6.8
 * 5 mM EGTA, pH 7.0
 * 2 mM MgCl2 ( final Concentration: 2 mM)
 * Yu-li Wang from Carnegie Mellon This is a pdf of how they do Rhodamine Tubulin
 * PEM buffer:
 * 0.1 M PIPES
 * 1.0 mM EGTA
 * 0.5 mM MgCl2
 * pH 6.9
 * Tedeschi of Italy
 * PEM buffer
 * 85 mM Pipes, pH 6.94
 * 10 mM EGTA
 * 1 mM MgCl2
 * Steve Gross of U Minnesota with Steve Block
 * 80 mM Pipes
 * 1 mM EGTA
 * 4 mM MgCl2
 * Stefan Diez (Germany)
 * BRB80 buffer
 * 80 mM potassium Pipes
 * 1 mM EGTA
 * 1 mM MgCl2
 * pH 6.9
 * Mathew Lang, Steve Block first author
 * Also used in Rosenfield, Block Paper
 * Also used in Guydosh Block Paper with 10μM of Taxol
 * Un-named buffer
 * 80 mM Pipes (pH6.9)
 * 50 mM potassium acetate
 * 4 mM MgCl2
 * 2 mM DTT
 * 1 mM EGTA
 * 7 mMTaxol
 * Andy mentions that he thinks these buffers are what Block used to get kinesin to stick to beads. It not be what we want, but if the whole "soup" (Beads + kinesin + buffer) is added to the microtubes mix and it doesnt mess things up, this stuff should be fine.
 * Valentine, Fordyce, Block
 * polymerization buffer for brain tubulin
 * 62 mM PIPES at pH 6.9
 * 0.8 mM EGTA
 * 3.7 mM MgCl2
 * 0.9 mM GTP
 * 10% v/v DMSO
 * stablilzation buffer
 * 80 mM PIPES at pH 6.9
 * 1.7 mM EGTA
 * 5.5 mM MgCl2
 * 1.2 mM GTP
 * 8.2 mM sodium azide
 * 20 μM taxol
 * 10% v/v DMSO)
 * Assay buffer based on PEM80
 * 80 mM PIPES at pH 6.9
 * 1 mM EGTA
 * 4 mM MgCl2


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