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One method to produce the powder involved snap freezing a lyophilized form of the material and then pulverizing it in a grinding mill.
Our measurements of TF by ELISA were done on lungs that had been lavaged (BAL) prior to snap freezing.
Aliquots of 1 ml each were taken from growing cells and quenched in chloramphenicol (200 µg/ml final concentration) prior to snap freezing in liquid nitrogen.
All lysates were prepared within 10 min prior to snap freezing.
Drosophila heads were extracted by snap freezing wild-type adult flies (age 1 4 days).
Another study compared the quality of DNA, protein, and RNA extraction from RNAlater and snap freezing (65).
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For adequately rapid fixation, snap-freezing is required.
The tissue was then rinsed in deionized water before snap-freezing in liquid N2 and ground into fine powder using a mortar and pestle.
Trummer, A., De Rop, C., Tiede, A., Ganser, A. & Eisert, R. Recovery and composition of microparticles after snap-freezing depends on thawing temperature.
This method takes advantage of the cell DNA replication machinery that remains active after snap-freezing.
We appreciate the good advice from Brita Robertson about snap-freezing.
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