PCR was performed with Q5 High Fidelity DNA polymerase (New England Biolabs), and dNTPs (Thermo Fisher Scientific). Primary PCR used primers “1st Step SWHEL forward” and “1st Step SWHEL reverse.” Conditions were 98°C for 3 minutes; then 98°C for 15 s, 62°C for 1 minute, and...
The PCR reaction mix contained 10×PCR buffer (PGC Scientific), 2.6% DMSO, 2 mM dNTPs, 2U Gene Choice TAQ polymerase (PGC Scientific), 200 nM each BAPANTI Forward and BAPANTI Reverse2 primers (Genosys) and 5 μl of diluted Ba a/g DNA in a 50 μl reaction volume. The primers ...
BecauseB. simplex30N-5 was originally isolated on an N-free medium, primers that targetednifHwere used in a colony PCR usingSinorhizobium(Ensifer)melilotistrain 1021 as the positive control. We did not find any evidence fornifHinB. simplex30N-5 despite the fact that we based our primer set...