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Afterwards, they usually find this procedure not necessary

If results look unusual, check your reconstitution entry

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Recombinants were selected on LB agar containing Kan incubated at 30 C overnight, and genes deletion in the BAC was confirmed by PCR amplification using Conf-KO BAC cobMK Mcan-Fw and P1 inv primers for the 5 end and P2 inv long and Conf-KO BAC cobMK Mcan-Rv for the 3 end