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Tissue preparation section of protocol booklet states the following:
10.3.1 Weigh the tissue and cut it into small pieces. Place tissue
pieces in a clean homogenizer.
10.3.2 Add 5 mL of 1X Pre-Extraction Buffer containing 5 μL of
DTT Solution per gram of tissue, and homogenize tissue
pieces (50-60 strokes).
10.3.3 Incubate on ice for 15 minutes and centrifuge for
10 minutes at 12,000 rpm at 4°C.
10.3.4 Remove the supernatant.
This does not mention PIC. Do we need to add this? Do they use stock solution of the pre extractio buffer, or do they use pre extraction buffer as prepared earlier in the protocol with DTT and PIC already added?
Asked on Jan 07 2016