I have tried the expression of a 20kD protein, cloned in pET3a in BL21 system. Bulk cultute is grown upto 0.5 OD at 600 nm and IPTG is added in 3 different conc. 0.1mM, 0.2mM and 0.3mM in 3 different sets and culture is collected after 1hr and 2hr induction. But I could not see the desired band in SDS-PAGE. Please give me suggestion.
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