[3dem] Recalcitrant viruses
Sacha De Carlo
sachadecarlo at yahoo.com
Wed Feb 27 17:32:05 PST 2008
whenever I have been working with polymerases (RNA pol II from Yeast or Human, or RNAP from Archaea) I always avoided glow-discharge, pre-soak the QF grids in chloroform for 30 min and let them dry in air prior to use. By following this very simple procedure I had RNA pols in the holes and not on carbon, on which they usually love to stick due to surface charges...
Maybe it can help you...
Eduardo Sanz-Garcia <esanzgar at chem.byu.edu> wrote:
I am working with a virus that likes a lot the carbon and doesn't want
to go to the holes.
Here are some pictures (the picture are not the best but it is possible
to see the effect):
I have tried glow discharged and no glow discharged grids.
I have also tried different type of buffers, virus concentration,
dialysis, and several applications of the virus in the grid.
Has anyone had a similar problem?
Does anyone have a solution for this problem?
Thank you very, very much
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3dem at ncmir.ucsd.edu
Sacha De Carlo, PhD
Molecular, Cell and Developmental Biology Department
Bio3D & Hoenger Labs, Campus Box 347
University of Colorado
Boulder, CO 80309
e-mail: sacha at bio3d.colorado.edu
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