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I am planning to isolate total proteins of gram positive bacteria. At final stage, I will resuspend the pellet in protein sample buffer that includes 40 mM Tris/HCl (pH 8) , 4 mM EDTA, 8% SDS,...
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I try to find sufficient parameters such as Hz of sonicator, sonication time, cycles and breaks...etc
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Taking samples during sonication and: 1)Visualize under microscope to see burst cells 2)Measure Absorbance at 280 for DNA/RNA release 3)Centrifuge and measure the protein concentration by...
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