Yeast breaking liquid nitrogen grinding method

asked:
what
reagents are needed to ask for detailed steps in the method of liquid nitrogen grinding of
yeast
broken walls
?
:
take 1L yeast, after induction, centrifugation (centrifugation) receiving body, weighing wet weight, purifying buffer suspended bacteria with protein( protein), weight (g): buffer volume (ml) . . . 1:1-1:2. Using a syringe to add the bacteria liquid into the liquid nitrogen, control the speed of addition, so that the bacteria liquid into the liquid nitrogen after the formation of small particles, too fast and easy to form blocks. Transfer the bacteria liquid nitrogen particles into the high-speed crusher (using the machine that breaks the herbs, RMB600), take care not to pour excess liquid nitrogen, about 2-3 million l/min broken for 1 minute, the bacteria particles should be powdered. Pour out the powder, melt in the 37-degree water bath, add liquid nitrogen with a syringe, repeat the above steps, repeat 3 times. High-speed (20,000 rpm, 4 degrees 30 minutes) centrifugation (centrifugation) to clear, such as more impurities, need to be more than a few times away, the target protein (protein) purification with E.coli.