Name Oligonucleotide sequence Annealing temperature Second primer Product size (bp) Amplification strategy A. Primers used in the initial multiplex PCR amplifications Ex1Fw AGG ACA AAG TCT CCT AAT CTT C      750    Ex1Rv CAA TGA TCT CTT GTA ACA TTG TAC       Ex2Fw  TAA CTG TAT CTC CTT TTC TAG C     533   Ex2Rv AGG TCA GTG ATA GAG AGT ATG       Ex4 Fw AAG TGT TTT ATA TCT AAT GTT TAC     304   Ex4 Rv CTT CAG GGC TTG GTC AAT ATA G        EX5Fw CAG AGC TTG GCA TAT TGT ATC     321   Ex5 Rv  GTA AAC ACA CAA CTA GTC AAT G       Ex7 Fw TGT TCC ATT TCT CTC CTT TTC C      272   Ex7 Rv GAA CAT GGA GTT GCA GTG TAG        Ex8 Fw CAT ATT AAC TTC CCT CAC TTC      174   Ex8 Rv  CTT GTA CCC TGA AAC ACA AAG        Ex9 Fw AGT AAC TTC TCC CTA TGT TTG      312   Ex9 Rv GAT GAC GGG TCA GAA GAA G        B. Allele specific Primers rs28399504, c.1A>G ACA AGA GGA GAA GGC TTC AA ACA AGA GGA GAA GGC TTC AG 61 Ex 1Rv  288 134 Duplexed with § c.395G>A, rs72552267 CTC CCT CAT GAC GCT GCG CTC CCT CAT GAC GCT GCA Ex 2Rv c.50T>C, rs55752064 CTC TCA TGT TTG CTT CTC CC CTC TCA TGT TTG CTT CTC CT 61  Ex 1Rv 239 298 Duplexed with § c.1297C>T, rs56337013 ATG TTT GTT ATT TTC AGG AAA AC ATG TTT GTT ATT TTC AGG AAA AT Ex9 Rv c.55A>C, rs17882687 CAG AGC TCT GTC TCC AGA T CAG AGC TCT GTC TCC AGA G 61 Ex1Fw Ex8Rv 554 122 Duplexed with § c.1228C>T, rs17879685 CCA GAG ATG TTT GAC CCT T  CCA GAG ATG TTT GAC CCT C  c.99C>T, rs17885098 AAT CAC TGG GAG AGG AGT G AAT CAC TGG GAG AGG AGT A 61 Ex1Fw 598 Duplexed with § c.636G>A, rs4986893 GTA AGC ACC CCC TGG GTA AGC ACC CCC TGA  Ex4Rv 109 c.276G>C, rs17878459 GCC TCT TCC AGA AAA CTC G GCC TCT TCC AGA AAA CTC C  63 Ex2Fw 147 488 Duplexed with ‡ c.449G>A, rs58973490 ACT CCT CCA CAA GGC AGC ACT CCT CCA CAA GGC AGT  c.431G>A, rs17884712 GAG GAG CAT TGA GGA CCG GAG GAG CAT TGA GGA CCA 61 Ex2Rv 98 With amplicons from PCR A+B‡ c.681G>A, rs4244285 CTA TCA TTG ATT ATT TCC CA CTA TCA TTG ATT ATT TCC CG 57 Ex5Rv 231 With amplicons from PCR A+B§ c.680C>T, rs6413438 CCA CTA TCA TTG ATT ATT TCC C  CCA CTA TCA TTG ATT ATT TCC T  61 Ex5Rv Ex7Fw 234 Duplexed with § c.991A>G, rs3758581 GCT CCG GTT TCT GCC AAT  GCT CCG GTT TCT GCC AAC  95 c.1251A>C, rs17886522 ACT TTC TGG ATG AAG GTG GC ACT TTC TGG ATG AAG GTG GA 61 Ex8Rv Ex9Fw 100 Duplexed with ‡ c.1473A>C, rs55640102 CAG ACC ATC TGT GCT TCT T  CAG ACC ATC TGT GCT TCT G  231

§Amplicons from A+B mixed and diluted to 1:25 with PCR buffer; ‡ Amplicons from A+B mixed and diluted to 1:50 with PCR buffer.

Table 1: Primers used in the initial multiplex PCR amplifications and allele specific PCR reactions with their cycling conditions.