Hi Cryo EM enthusiasts.

I have some 2nm carbon covered QF grids and want a bit of a sweep to see what experience people have with using these before I blindly start guesstimating gridding conditions. Some specific questions:

- What protein concentration do people generally work with relative to negative stain concentration or normal ice concentration. If I did neg stain on a protein at say 50 ug/ml and it looked nicely dispersed I would probably start looking at the protein at about 1 mg/ml. So for carbon coated QFs would 150ug/ml be the ballpark to start?

- Do people glow discharge the grids briefly before use?

- How does blot time compare to say a normalish 4 second blot. Do people find they go shorter or longer?

thanks

Rich