[3dem] Ice thickness

[Bowman, Valorie D]

And bracket them with as many multiples of blot times and offsets as you have sample and patience to allow!


Cheers,

Valorie

Valorie Bowman

vdb at purdue.edu
765-494-5643
EM Facility Laboratory Manager/
Senior Research Electron Microscopist
Purdue Cryo EM Facility
Purdue University
________________________________
From: 3dem <3dem-bounces at ncmir.ucsd.edu> on behalf of David Michael Belnap <David.Belnap at utah.edu>
Sent: Monday, February 26, 2018 11:57 AM
To: Sanchaita Das
Cc: 3dem Scripps
Subject: Re: [3dem] Ice thickness

Sanchaita,

"There is no royal road to [success in cryo-EM]."  :-)

I would start with settings that have been successful with other samples on that Vitrobot.

David


On Feb 26, 2018, at 9:09 AM, Sacha De Carlo <sachadecarlo at yahoo.com<mailto:sachadecarlo at yahoo.com>> wrote:

Unfortunately the molecular weight of the complex alone is useless to determine "proper" settings for the vitrobot. Buffer composition, sample concentration, etc would be required to give you an educated guess ...

Cheers
Sacha


Sent from Yahoo Mail for iPhone<https://yho.com/footer0>


On Monday, February 26, 2018, 5:05 PM, Sanchaita Das <sanchaita06 at gmail.com<mailto:sanchaita06 at gmail.com>> wrote:

Dear all,
Does anyone have suggestions about what vitrobot settings may work for getting good single particles for a protein that is aproox 300KDa?
I know its trial and error but if something might work for smaller proteins would love suggestions.
Best,
Sanchaita


I think 99  times and I find nothing.
I stop thinking,
swim in silence,
and the truth comes to me.

- Albert Einstein -

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