Tuesday, April 8, 2014

bio125, protein extraction

I spent 50 minutes to explain the protocol on board. Students were given spundown cells in eppendorf tubes, and begin with adding 160 lysis buffer.  The protein extraction took 1 hours and finished around 3:10pm. I used the rest of the time for course evaluation.

Video: http://youtu.be/piC1Koyu1No

yeast cell wall, membrane, nuclear envelope

Nuclear and cytosol

beta mecaptal ethanol


Protocol on board: 

After adding blue ESB to the protein pellet, the TCA will turn ESB into yellow. Use Tris to adjust the color back to blue.

Add DSB to pellet
Changed to yellow. 
Adjusted to blue. 

MSH2 expression, cell cycle, log-phase
hMSH2 has no putative NLS signal, but MSH6 have 3.
co-import is supported.

MSH2 deficient,

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