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Cell pellets were resuspended in 0.3 ml small-scale lysis buffer (20 mM potassium phosphate, 200 mM NaCl, pH 7.4, 1 mg ml 1 lysozyme, 0.75 mg ml 1 polymyxin B, 0.01 mg ml 1 DNase I) per well and incubated at 30 C, 300 rpm, 5 cm shaking diameter for 30 min

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HCG is injected only ONCE a day, typically in the morning