Table 3.

Characteristics of the 36 novel EST-SSR markers in R. rex.

Primer	Forward primer (5″–3″)	Reverse primer (5″–3″)	Tm (°C)	SSRs	Number of alleles	Ho	He	PIC	HWE	Fst	r
Rho-1	ACCGAGTCACAGCACTCCTT	CACATTCATCCTCCCCAATC	52	(CGC)5	3	0.050	0.165	0.412	0.0003***	0.676	0.331
Rho-4	TTGACGAACTGCACCAACTC	TTCAGTCACAAACCTGCATCA	55	(TG)6	3	0.083	0.153	0.166	0.0001***	0.323	0.231
Rho-10	GGGAGAGAGAGGTCCTACCG	CTGCCCTTGTTTGACGATTT	60	(GA)7	11	0.433	0.685	0.870	0.0000***	0.221	0.265
Rho-11	GAGGACGAGGGTGGTACAAA	AAGCCATGGAGTTGATACGG	60	(GCA)7	3	0.350	0.299	0.338	0.0000***	0.222	0.135
Rho-13	AGCCCTTCCTCTGTCTCCTC	TTCGAATGGATCAAATGGGT	58	(CTC)5	4	0.200	0.160	0.208	0.9998ns	0.141	0
Rho-14	GAGATTCTCAACCCAACCCA	TCCAAACAGACATCCGATCA	51	(GA)6	5	0.263	0.342	0.490	0.0000***	0.390	0.226
Rho-15	CAATCAAGGGGCTACCATGT	CGAAAGTTGGTGGTATCCGT	57	(AGG)6	5	0.700	0.580	0.618	0.1278ns	0.134	0.241
Rho-17	AGTGGACAGTGAGGTCACCC	TCGGATGAATTGCGTTGTAA	60	(TGT)5	4	0.588	0.391	0.520	0.8759ns	0.364	0.578
Rho-20	TCCTGGGTCCATAACACACC	GGTCACGTGTCTGAGCGTAA	60	(AC)9	2	0.200	0.120	0.352	0.0325*	0.736	0.485
Rho-26	CCTGAATCCATCCTGTCCTG	GCTGAGGGATCACCAGACAT	56	(GA)8	9	0.700	0.583	0.798	0.0024*	0.303	0.176
Rho-27	GGGGTAATACCGGAGGGTAA	GTTCCCTGAAGACATGGTGG	56	(GA)7	3	0.200	0.295	0.406	0.0012*	0.312	0.391
Rho-30	GGAAGTTTCGGCAGCAGTAG	CCTTCTCCCAACTCCCTTTC	56	(GGT)5	7	0.650	0.498	0.564	0.2730ns	0.169	0.11
Rho-33	CCACCCTTCCCTTATCCTTG	GAGAAGTGTGGCTTTGAGGG	56	(CT)7	9	0.450	0.571	0.737	0.5345ns	0.237	0.224
Rho-34	AGTGGCCTTAGGGGAAAGAA	CAACCCTTACCCACCACATC	56	(GGC)5	2	0.050	0.045	0.053	1.0000ns	0.077	0.972
Rho-35	TGAATCCACCACAACAAGGA	ACTCCCCTCTCGGAAATTGT	55	(TA)8	4	0.333	0.334	0.473	0.1908ns	0.392	0.161
Rho-37	CATGGAGAAGACCCACTGGT	ACCCACGCATTAACTTCAGG	56	(CT)8	9	0.450	0.698	0.820	0.0000***	0.169	0.324
Rho-39	GTGGCTCAAAATACAGGGGA	CAGATGAAGGCGATGTGAGA	58	(GCT)5	5	0.150	0.355	0.368	0.0129*	0.123	0.209
Rho-45	CTATGGCGGGCCTATCTGT	CATACGAAGGACGAGGTGGT	60	(TCC)6	8	0.400	0.670	0.700	0.0000***	0.092	0.238
Rho-48	CTGCGTCTTTGGGTTTCTTC	CAATCCAACCCACCATTTTC	55	(GAG)5	5	0.113	0.313	0.333	0.0000***	0.167	0.238
Rho-51	AAATATGTTCACCCCCACCA	GCCTGGACTGTTGGAATGTT	58	(CAC)5	5	0.163	0.262	0.271	0.0058*	0.097	0.175
Rho-53	AACCAGTACAGGACGCCAAG	CTCCCGAGAAGATCAAGCAG	56	(GCG)5	3	0.050	0.345	0.345	0.0001**	0.161	0.481
Rho-54	GAAATACGGAAACGGACGAA	CCTCTCTCTCTCCGCACATT	52	(TGTA)5	3	0.392	0.332	0.517	0.1424ns	0.435	0.26
Rho-56	CCTCCTCTCGCATCATTCTC	CACTGCCATCTCTCACTCCA	56	(TCG)5	4	0.213	0.207	0.237	0.0001**	0.169	0.11
Rho-59	AAACCTCTCGCTCTCTTCCC	GGTGTCGGTCTTCATGGTTT	56	(CT)7	9	0.725	0.623	0.821	0.1762ns	0.260	0.096
Rho-62	ATATGTTGCGCGGGAGAAT	TCTCGAAGGCAAAACAGCTT	54	(CT)6	5	0.250	0.345	0.365	0.0000***	0.101	0.195
Rho-67	GGTGGATCAGAAGGGACTGA	ACATGAAGATCATGGGCGAT	55	(CTT)5	8	0.375	0.501	0.579	0.0000***	0.169	0.196
Rho-69	CGAATCCTCCATCAAAGCAT	ATGCAAAACTGTGACCTCCC	57	(TTC)5	5	0.100	0.355	0.651	0.0000***	0.459	0.425
Rho-70	GGCTGTGAGGGAGTCAAAGA	TCTCCATTGTCGAAACCTCC	56	(GA)6	7	0.450	0.383	0.410	0.0246*	0.162	0
Rho-74	ATAACGCGCAAACTAGCGTT	ATGAGGAGGAGCGCACTTTA	58	(CTG)5	6	0.400	0.455	0.671	0.0001**	0.366	0.502
Rho-75	GTAAATGGGCCCGTATTCCT	CTCCATTGAGAAACCCTCCA	54	(TTG)6	4	0.150	0.120	0.154	0.9999ns	0.158	0
Rho-79	TGGTTCTGTTCTCTGGCCTC	TTCCAGGATAGTGCTCCTGC	58	(TC)7	9	0.400	0.415	0.684	0.0000***	0.428	0.22
Rho-81	ATGGATCGTTCTGGACGAAG	AAGGCCACTAGAAGAAGCCC	60	(CT)8	10	0.613	0.601	0.801	0.0011**	0.275	0.232
Rho-84	TGACGGACTTGTGCTGGATA	GAGAAAAGGGAAGAAGGACACA	58	(TC)7	2	0.050	0.310	0.372	0.0002***	0.373	0.642
Rho-93	TGCAGAGTAAAACCCTGCTTG	TAAAGTTGAGGCGGCAAAGT	58	(CA)7	3	0.050	0.085	0.099	0.0000***	0.117	0.153
Rho-95	GGGGTAGGGGGATACTTTGA	GTCGACGACTTTGGTCCAGT	55	(GCT)6	8	0.450	0.395	0.509	0.0001***	0.249	0.084
Rho-97	TTTGCGGTGGTGTCTGAATA	AAATCCAATGATCCATCCCA	55	(GA)7	5	0.425	0.404	0.631	0.0016**	0.412	0.159
Mean					5.472	0.323	0.372	0.482		0.268	0.263

ns, non-significance;

^*p < 0.05, significant difference;

^**p < 0.01, most significant difference;

^***P < 0.001, most significant difference; r, null allele frequency.