Pooled sample consisting of samples: CystA 1009 S1|CystA 1033 S1|CystA 1049 S1|CystA 1081 S1|CystA 1026 S1|CystA 1042 S1|CystA 1050 S1|CystA 1019 S1|CystA 1027 S1|CystA 1067 S1|CystA 1004 S1|CystA 1012 S1|CystA 1028 S1|CystA 1007 S1|CystA 1015 S1|CystA 1008 S1|CystA 1024 S1|CystA 1048 S1
Primers were used to amplify DNA in 25-uL multiplex polymerase chain reactions (PCRs) as described previously 15.  For each sample, three multiplex PCRs were performed, with each multiplex PCR containing 22 to 50 primer pairs.  Reactions were purified with AMPure XP beads (Beckman Coulter) and eluted in 100 uL of Buffer EB (Qiagen, cat. no. 19086).  0.25% of the purified PCR products were then amplified in a second round of PCR, as described in Springer et al.15.  The PCR products were purified with AMPure and used for sequencing on a MiSeq or HiSeq instrument