User:Linh N Le/Notebook/2009/06/22: Difference between revisions

From OpenWetWare
< User:Linh N Le‎ | Notebook‎ | 2009‎ | 06
Jump to navigationJump to search
Line 49: Line 49:
**I need 15.99884 g for .4L of water (4 deca liters)
**I need 15.99884 g for .4L of water (4 deca liters)
**divide by 10, 1.599884g for a 40ml
**divide by 10, 1.599884g for a 40ml
===Making the Buffer==
===Making the Buffer===
*We measured out the following
*We measured out the following
**1mM EGTA (0.076g)
**1mM EGTA (0.076g)
Line 57: Line 57:
*We titrated it with NaOH to raise the Ph to 6.89
*We titrated it with NaOH to raise the Ph to 6.89
**Used about 26ml of NaOH to get there
**Used about 26ml of NaOH to get there
 
*Added an entry to the Kochlab inventory page, under the VWR fridge category
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
|}
|}

Revision as of 16:08, 22 June 2009

Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

To Do

  • Andy is back! He wants to try the kinesin again, maybe his expertise will allow us to succeed
  • Make BRB80

Tubulin

  • Stumbled upon a site that details how they get tubulin from cow's brains

BRB80

  • Since Andy and Koch are the only people in the lab that know how to make a buffer (I actually dont know if Ant can), we waited til Andy came back to try to make BRB80
    • Not to mention the fact that we are also running out of the stuff we got from Cytoskeleton (new shipment will be in this week)
  • Andy is looking over our new fancy Ph meter
    • He is starting the calibration process
  • BRB stands for Britton-Robinson Buffer
    • It is a "universal" buffer whose ph changes linearly with the amount of alkali that is added to it
    • It consists of:
      • .04M H3BO3 (Boric Acid)
      • .04M H3PO4 (Phosphoric Acid)
      • .04M CH3COOH (Acetic Acid)
  • The stuff we use is also known as PEM
    • 0.1 M PIPES (pH 6.95)
      • Piperazinediethanesulfonic acid C8H18N2O6S2
    • 2 mM EGTA
      • ethylene glycol tetraacetic acid C14H24N2O10
    • 1 mM MgSO4
      • Magnesium Sulfate
  • Also can make it as
    • 100 mM Pipes,pH 6.9
    • 1 mM MgCl2 (magnesium chloride)
    • 1 mM EGTA

Ph Titrations

  • Recalling what little i remember from freshman chemistry, Andy has instructed me to create 40uL of a "normal" base of NaoH
    • Normal meaning 1M soln
  • So, we look up the molar mass of NaOH
    • MM 39.9971 g/mol
  • If I want a 1 mol soln, i need 39.9971 g in 1L
    • Means I need 3.9971g in a dL
    • Means I need .39971g in a ml
    • Means I need .039971g in a ul
      • Meaning I need 1.598g in a 40ul soln
  • Thats wrong, new train of thought
  • Lets try to make .4L of 1M soln
    • I need 15.99884 g for .4L of water (4 deca liters)
    • divide by 10, 1.599884g for a 40ml

Making the Buffer

  • We measured out the following
    • 1mM EGTA (0.076g)
    • 80mM PIPES (4.83g)
    • 1mM MgCl2 (.2mL)
  • For a 200mL stock of the Buffer (which Andy hates naming, but I will name KLB)
  • We titrated it with NaOH to raise the Ph to 6.89
    • Used about 26ml of NaOH to get there
  • Added an entry to the Kochlab inventory page, under the VWR fridge category
Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Linh_N_Le/Notebook/2009/06/22&oldid=317966"