Table 5

Gene	Intron size (bp)	Input sequence (bp)	Primer	Primer Sequence	TM (°C)	GC (%)	Product size (bp)

1	136	601	F	AGTCCACATCTCTTTGCG	54.6	50	220
			R	CCAAGTCAATGACAGAGTCC	55.5	50
2	95	251	F	CACCAACGTAGGGCTACAC	56.6	58	297
			R	CTTTAAAGGTGAGGGCGT	55.4	50
3	1261	350	F	CATCAGCATACTCTCCTTCC	54.8	50	282
			R	TAACCGTAAGGTGTCTCCAC	55.2	50
4	4899	700	F	TGGACTAGCCAATGATGG	55.3	50	234
			R	AACACTAAAACCCCCTCG	55	50
5	88	280	F	AGCCTCTGCTAGTGCCTCTG	60	60.0	132
			R	AGCCTGCAGAGCTAGGGTTT	60.5	55
6	6122	950	F	GGGCTTGGCTTAAATCTTCC	60	50	240
			R	ATCAAATGGCTTCCATCTCG	60	45
7	81	350	F	CACCCCAAAATAACCCAAAA	59.5	40	121
			R	TGCTAACGAAGAACGCTTGA	59.8	45
8	74	420	F	TTCACAACCCTATTGTGGCA	60	45	286
			R	GTCCATGATTGCCTCAGCTT	60.2	50
9	274	350	F	CTCCAATGGTCTTGGATCGT	60	50	191
			R	AAGGAAGTACTGAGTGCGGC	59.5	55
10	483	350	F	TGTGTGCTTGTGTTTGTCTCTT	58.5	41	218
			R	CAAACACACTGAGCCACAGAC	59.4	52.4
11	476	230	F	GGGCTGATCCAAGAGACAAA	60.2	50	200
			R	ATCGACATCCCTTCCAACAA	60.3	45
12	73	230	F	TATGGCTACCCATCTGCCTC	60.1	55	189
			R	TCATTCCCACCATCTTGGAT	60.1	45
13	1689	420	F	ACACAACAATTGGTCTGCCA	60	45	227
			R	GAGGAATCTTAGGGAACCGC	60	55
14	252	280	F	TGAGCATACATATTCTGCAACAAA	59.7	33.3	152
			R	GGGTTAGTTGGACTTTGGGG	60.6	55
15	82	280	F	GTACCAGGCAGAAGGGAACA	60.1	55.0	218
			R	AGTGTTTGGGAACCTGAACG	60	50

Product size corresponds to the expected band size in a gel- electrophoresis assay, following successful amplifying of each predicted genes. Primer F is forward primer and R is reverse prime for the corresponding predicted gene in soybean. Primers highlighted with GC content less than 45% need to be extended as explained in the discussion section

Table 5: The predicted genes, with corresponding introns sizes, and designed primer pairs, with melting temperature (TM), GC bases content and product size genes that are linked to marker Satt460 and the putative Rpp3 locus.