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steriods The position of the cointegrate junctions was confirmed and the orientation determined using primer pairs RH1471/RH1452 and RH1472/RH1451 to link the R388 backbone across each IS26 into the pRMH762 derived region (Fig.5). PCR products of the expected size (1,464bp and 1,042bp, respectively) confirmed that all of the tested cointegrates shared the same boundaries between pRMH762 and R388::IS26, and the IS26s were always in the direct orientation. The sequences of the PCR amplicons showed that, in every case, the direct repeats flanking the single IS26 in R388::IS26 were now located next to the left and right ends of the IS26 flanked pRMH762 and the direct repeats surrounding the IS26 in pRMH762 were at the internal boundaries (Fig.5).. steriods

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