Table 1 Primer sequence and polymerase chain reaction (PCR) conditions for SPRED2, SPRED3, SPRY1, SPRY2, SPRY3 and SPRY4
GeneExonPCR fragmentsForward primer (5′–3′)Reverse primer (5′–3′)PCR temperature (°C)PCR buffer
Spred 211aaggaagctttttgcactccagatagggtttgggggaagg65Failsafe D
1bcgtccccttcccaaatctacgccaagttcacccaataa61Failsafe D
2tgccctgtgcttcttcttctcccacgagttgtatgtctgc6110× PCR buffer
3ttggtagttgccaccagtgatactgacctggtcccaaacc6110× PCR buffer
4tgcaaaaagaatggaatgtggacagcaacagcaaacactgc6410× PCR buffer
5ttttgggaagagcagcaaattaaaggcaggagcaggagag6110× PCR buffer
66aggagaaggtggggtgtttttctttggggtcctcgccta6110× PCR buffer
6bgaggtccccaagcatgactaaaggcaataagagccatcca61Failsafe D
6cggaccccgagggagactataagagacgagttcccctgtg6110× PCR buffer
Spred 31tgtgttgtccctgtccttccacctgacctctgctccacat61Failsafe D
2+3Exon 2+3 togetheratggacgagacgtcacagggcctgctacacagtaggtgct68Failsafe E
4gattctgagaagcgggaagaacgtgggactggaggagatt6110× PCR buffer
1ccctcctcctaccacagagagaaaagcccattttcaagca6010× PCR buffer
Spry 122accccagcatcattgtaatcccgaggagcaggtcttttcac6010× PCR buffer
2btgctattttgtccttagaccagaggaacccttcaagtcatcca6010× PCR buffer
2cccagtccctggtcataggtcctaagcacatgcaggttcca6010× PCR buffer
2daccatcctgtttggcctgtaggtgggaaacagggaaaaat6010× PCR buffer
1acttggcgatctggctttagcaggaggggaagagccaaac6010× PCR buffer
Spry 222.1attcaggtgggtggagagtgagctgaccgtgcttatggat6010× PCR buffer
2.2ctccactcagcacaaacacgtggctggaaggtaacaccat6010× PCR buffer
2.3gacaacccatgttcttgcaggcctaggagtgtctgtgttgc6010× PCR buffer
2.4cggtgaaatgctttttgtccttttgccttacagaaagccttt6010× PCR buffer
2.5tttgcaactgtgaaaaacaagtgataaaagaagacaaaacagctcagt60Fail safe C buffer
11aatgtgccctgaaatccatgtactgcgggggagagaagtag6010× PCR buffer
Spry 31btgtaaacaggccctctccagtcagcttctcccttcagagc6010× PCR buffer
1ccttcaggccaatccatcatccatcatcagtggagcagtgg6010× PCR buffer
1dgctgagagcctcctcgattaggggactcgaaggagaaaag6010× PCR buffer
1tgtgcccacccattcatagtgagggaaggaacactgagg6010× PCR buffer
Spry 42taagtcaaggggagggtgtgcaggatatcaaaatccccagaa6010× PCR buffer
33aggctcccaaggacctgatggtgacatcctggtcacagc6010× PCR buffer
3btagcctggccctgaccacagcacaggcactcctggtt6010× PCR buffer
3caaatgcaaggagtgtgcatccaggcaggtagcagagcag6010× PCR buffer
3dggtgctctctccgtggtgttcttagatgtcagaagagaacca6010× PCR buffer