Pooled sample consisting of samples: CystA 1102 S1|CystA 1103 S1|CystA 1104 S1|CystA 1105 S1|CystA 1107 S1|CystA 1108 S1|CystA 1109 S1|CystA 1120 S1|CystA 1121 S1|CystA 1122 S1|CystA 1123 S1|CystA 1125 S1|CystA 1126 S1|CystA 1127 S1|CystA 1128 S1|CystA 1129 S1|CystA 1130 S1|CystA 1131 S1|CystA 1132 S1|CystA 1133 S1|CystA 1134 S1|CystA 1136 S1|CystA 1137 S1|CystA 1138 S1|CystA 1139 S1|CystA 1140 S1|CystA 1141 S1|CystA 1142 S1|CystA 1143 S1|CystA 1144 S1|CystA 1145 S1|CystA 1146 S1|CystA 1147 S1|CystA 1148 S1|CystA 1149 S1|CystA 1150 S1|CystA 1151 S1|CystA 1152 S1|CystA 1153 S1|CystA 1154 S1|CystA 1155 S1|CystA 1156 S1|CystA 1157 S1|CystA 1158 S1|CystA 1159 S1|CystA 1160 S1|CystA 1161 S1|CystA 1162 S1|CystA 1163 S1|CystA 1164 S1|CystA 1165 S1|CystA 1166 S1|CystA 1167 S1|CystA 1168 S1|CystA 1169 S1|CystA 1170 S1|CystA 1171 S1|CystA 1173 S1|CystA 1175 S1|CystA 1176 S1|CystA 1177 S1|CystA 1178 S1|CystA 1180 S1|CystA 1181 S1|CystA 1183 S1|CystA 1185 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