Scaling up 100nl drops is problematic. What I understand is that it is not only the different equilibration conditions, but primarily the amount of protein that gets absorbed on the surface is relatively higher for small drops. There were some empirical formula for scaling up (i.e. how much you need to increase the protein concentration, but I am afraid you would have to re-screen anyway.
On Mon, 2012-03-19 at 15:31 -0500, Rajesh kumar wrote: > Dear All, > > > I have few papers in hand which explain me about microseeding, matrix > microseeding, and cross seeding. > I have also read few earlier threads and some more literature in > google. > Using Phoenix robot, I did a matrix micro-seeding and matrix cross > seeding. I have few hits with this. > In 96 well I used 100+100+50 nL and 200+200+50 nl (protein+screen > +seed) in separate expts. > I have hard time to plan to translate this 96 sitting drop well plate > to 24 well plate to refine the conditions to get better crystals. only > 1-2 hits are small crystals and they are tiny. > > > I wonder in 24 well plate, if I should do- > 1) for Example 500+500+50nl (I am sure I cant add less that 500 > nL precisely) > 2) to a drop of 500+500 nL do microseeding/streaking with a hair > > > I appreciate if you could advise and share some practical ways to > further my experiment. > > > Thanks in advance > Regards, > Rajesh -- Edwin Pozharski, PhD, Assistant Professor University of Maryland, Baltimore ---------------------------------------------- When the Way is forgotten duty and justice appear; Then knowledge and wisdom are born along with hypocrisy. When harmonious relationships dissolve then respect and devotion arise; When a nation falls to chaos then loyalty and patriotism are born. ------------------------------ / Lao Tse /
