Assessing meiofaunal variation among individuals utilising morphological and molecular approaches: an example using the Tardigrada

Table 1 Primers used in each reaction combination

		Forward	Reverse
18S
Amp 1		SSU01_F	SSU82_R_short
		AACCTGGTTGATCCTGCCAGT	TGATCCTTCTGCAGGTTCACC
Amp 2
	Fragment 1	SSU01_F	SSU26_R
		AACCTGGTTGATCCTGCCAGT	CATTCTTGGCAAATGCTTTCG
	Fragment 2	SSU22_F	SSU13_R
		TCCAAGGAAGGCAGCAGGC	GGGCATCACAGACCTGTTA
	Fragment 3	SSU26_F	SSU82_R
		CGAAAGCATTTGCCAAGAATG	TGATCCTTCTGCAGGTTCACCTAC
CO1
Amp 1		LCO_1490	mtD9_2206
		GGTCAACAAATCATAAAGATATTGG	CCCGGTAAAATTAAAATATAAACTTC
Amp 2		LCO_1490	HCO_2198
		GGTCAACAAATCATAAAGATATTGG	TAAACTTCAGGGTGACCAAAAAATCA
Wingless
Amp 1		Wg1s_F	Wg2n_R
		GARTGYAARTGYCAYGG	ACYTCRCARCACCARTG
Amp 2		Wg1a_F	Wgs_R
		GARTGYAARTGYCAYGGYATGTCTGG	ACYTCRCARCACCARTGRAA
Alpha Spectrin
Amp 1		Aspec11_F	Aspec15_R
		TGGATHMGNGARAARGA	AARTCRTCRAAYTTYTTYTG
Amp 2		Aspec11_F	Aspec12_R
		TGGATHMGNGARAARGA	ACYTCNACYTTCCACCARTC
Elongation Factor 1alpha
Amp 1		237_F	1450_R
		CGGYCAYTTGATCTACAAATGC	TGTCRCGCACAGCGAAACKACC
Amp 2		277_F	1221L_R
		ACSATYGAGAAGTTCGAGAAG	GGRTGRTTMARIACRATMACCTG

Each sequence corresponds to one of two oligo-nucleotide primers used for each specific reaction. In addition, all forward primers were tailed with M13_REV (CAGGAAACAGCTATGACC) and all reverse primers were tailed with M13 – 21 (TGTAAAACGACGGCCAGT). Primers for 18S were adapted from [55]. LCO/HCO primers for CO1 were designed by [56]. mtD9 was designed by [57]. Wingless and Alpha Spectrin primers were sourced from [58]. Elongation Factor 1 alpha primers were designed from GenBank alignments of onychophora, tardigrade, drosophila and artemia.

ISSN: 1472-6785