Table 2. 2-Plex PCR primers used in Fig 3B.

Lane ID	Name	Multiplex Primer Mix				F2_R1 Short Amplicon
		Amplicon1		Amplicon2		Loop Length(nt); Stem Oligo; Stem length(nt)
1	SinglePlex Control mixa					no stem loop
2	tStem_t1	t2_F1	t1_R1	t1_F2	t2_R2	195; t1_only; 20
3	tStem_4nt-t1	t2_F1	4nt-t1_R1	4nt-t1_F2	t2_R2	195; t1+4nt; 24
4	tStem_8nt-t1	t2_F1	8nt-t1_R1	8nt-t1_F2	t2_R2	195; t1+8nt; 28
5	tStem_12nt-t1	t2_F1	12nt-t1_R1	12nt-t1_F2	t2_R2	195; t1+12nt; 32
6	tStem_16nt-t1	t2_F1	16nt-t1_R1	16nt-t1_F2	t2_R2	195; t1+16nt; 36
Marker	1Kb Plus Ladder					Not applicable
7	sStem_t1-F2^4	t2_F1	t1-F2^4_R1	t1_F2	t2_R2	195; t1+4nt of F2; 24
8	sStem_t1-F2^8	t2_F1	t1-F2^8_R1	t1_F2	t2_R2	195; t1+8nt of F2; 28
9	sStem_t1-F2^12	t2_F1	t1-F2^12_R1	t1_F2	t2_R2	195; t1+12nt of F2; 32
10	sStem_t1-F2^16	t2_F1	t1-F2^16_R1	t1_F2	t2_R2	195; t1+16nt of F2; 36

The reactions in the gel electrophoresis in Fig 3B contains tStem primers from Table 1 in Lanes 2–6 and sStem primers in lanes 7–10.

^aContains each singleplex product from separate amplifications mixed at an equal volume ratio.