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A final rule entitled Medicare Clinical Diagnostic Laboratory Tests Payment System (CLFS final rule), which appeared in the Federal Register on June 23, 2016 (81 FR 41036), implemented section 1834A of the Act at 42 CFR part 414, subpart G

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Assuming you followed the above advice and used an appropriate binding buffer and your protein isnt huge, then a likely reason for this is that the expression yield is much lower than you expected

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