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. 2017 Jul 12;12(7):e0181062. doi: 10.1371/journal.pone.0181062

Fig 3. Tag-only stem and SLIMamp 2-plex PCR.

Fig 3

A) Four amplicons are produced during the amplification of two overlapping amplicons. B) Gel electrophoresis of two oligo sets (tStem and sStem) that form stem-loop structures. Lane M: 1Kb Plus ladder. Lane 1 is a singleplex control where Amplicon 1 and 2 were amplified separately and combined for electrophoresis. Lanes 2–6: mPCR with tag-only stem (tStem). The F2 and R1 primers form a stem, but the entire length of F2 for the overlapping amplicon is available for annealing in the loop of the stem-loop structure. Lanes 7–10: mPCR with SLIMamp primers. The stem length is similar to lanes 3–6, but the stem is formed with F2, creating competition for annealing with the primers. Amplicon 3 was greatly inhibited in the SLIMamp reactions but not in any of the tStem reactions.