Table 2. Details of primers: plant species targeted, primer sequences (forward- followed by reverse primer), expected amplicon length, concentration of each primer (µM), optimized annealing temperatures (°C), MgCl₂ concentration (mM), and affiliation to a multiplex assay.

Target taxa	Primer name	Primer sequence (5′-3′)	Size (bp)	Conc. (µM)	°C	MgCl2(mM)	Multiplex assay
Fagopyrum esculentum	Fag-sp-S519	gaaaacgaaaggaaaggttcat	380	0.2	56	4	FLPS
	Fag-sp-A523	caggattacccgttttttga
Lolium perenne	Lol-per-S528	gcatttttctatatagaatggat	254	0.4
	Lol-per-A535	tgactctatgttctccttagtt
Phaseolus coccineus	Pha-sp-S525	atcctttcacaaaaattccag	235	0.2
	Pha-sp-A531	tggatcagttcttcaagggt
Sinapis alba	Sin-alb-S534	attcactaaactactagatcgt	203	0.4
	Sin-alb-A542	catgaaatcaaaattcgaaagtc
Lupinus angustifolius	Lup-sp-S522	gaatccattcaacagttctg	244	0.2	53	4	LF
	Lup-sp-A527	gaacttttctttgtttttgcg
Fagopyrum esculentum	Fag-sp-S519	gaaaacgaaaggaaaggttcat	206	0.2
	Fag-sp-A524	tattaccctttcataccgcat
Taraxacum officinale	Tar-sp-S546	cggttcaaaactcctttatg	194	0.2	58	4	TAT
	Tar-sp-A554	ttcctcatgtctcatcctt
Achillea millefolium	Ach-sp-S547	gcggttcaaaattccttatac	222	0.2
	Ach-sp-A556	agggtattacaaagactcg
Trifolium repens	Tri-sp-S550	cagtaggaaaggaatcgttct	172	0.2
	Tri-sp-A558	aatctttcatttgtgatagaaaag
Trifolium pratense	Tri-sp-S550	cagtaggaaaggaatcgttct	151	0.2
	Tri-sp-A558	aatctttcatttgtgatagaaaag
Triticum aestivum	Tri-aes-S536	gctattaactagttctaaatttgaagtta	306	0.5	54	4	TZ
	Tri-aes-A545	cctcccgtcttacttttttat
Zea mays	Zea-may-S510	atttgatcattatatacatttttgagat	181	0.2
	Zea-may-A539	tccttccttttttagagtattcc
Plantago spp.	Pla-sp-S557	atctattttctagctatcctacc	116	0.5	61.5	4
	Pla-sp-A565	cgcatgtgataagagaaagtc
Apiaceae	Api-gen-S561	aatgaccgtctttgaccaaa	198/199	0.5	62	3
	Api-gen-A569	attctcattcccgatatcgc
Poaceae	Poa-gen-S541	gctttctcattctactctttc	187–293	0.2	56	3
	Poa-gen-A551	cttttcttgtgcatcatcctag

The genus- and family-specific primers were run in singleplex reactions.