Hello Georg, You can melt the agarose in a glass container using a microwave oven. Then connect your capillary tube to small syringe, using a plastic tubing or adapter. Use some parafilm if the tubing diameters do not fit exactly, there should be no leaks. Then slowly pull the plunger to take ~100ul of agarose and keep pulling outside the liquid, until the plug is more or less in the center of the tube, leaving room for the precipitant and protein on each side.
But most importantly, bear in mind that the system in the paper you cite works because the mother liquor components can diffuse readily through the agarose plug (MPD, NaAc, AmSO4, CaCl2). This might not be the case if your precipitant has high molecular weight PEG or the like, and your crystals need lower temperature to grow. Good luck, Javier On Mon, Jan 5, 2015 at 11:25 AM, Georg Mlynek <[email protected]> wrote: > Dear Colleagues, > > After reading a few papers about growing suitable crystals for neutron > diffraction. I will do capillary counterdiffusion with an agarose plug > between mother liquor and protein solution like described in > http://www.ncbi.nlm.nih.gov/pubmed/23192028. > > However I looked quite some time now, to find how to put the 100 ul 1% > low-melting agarose plug in the middle of the capillary, but was not > successful. Does one use a gel loading tip or is there a better way to do > this? > > Thanks in advance for any tips and tricks, > > best regards, Georg. > -- Javier M. Gonzalez, PhD. Protein Crystallography Station Bioscience Division Bioenergy and Biome Sciences Group (B-11) Los Alamos National Laboratory TA-3, Building 4200, Room 202B Mailstop T007 Los Alamos, NM 87545 Phone: +1 (505) 667-9376 LinkedIn <http://www.linkedin.com/pub/javier-gonz%C3%A1lez/22/7b/83a> Email <[email protected]>
