Pooled sample consisting of samples: Cyst 549 S2|Cyst 553 S2|Cyst 554 S2|Cyst 565 S2|Cyst 568 S2|CystA 1201 S1|CystA 1202 S1|CystA 1203 S1|CystA 1204 S1|CystA 1205 S1|CystA 1206 S1|CystA 1207 S1|CystA 1208 S1|CystA 1209 S1|CystA 1210 S1|CystA 1211 S1|CystA 1212 S1|CystA 1213 S1|CystA 1214 S1|CystA 1215 S1|CystA 1216 S1|CystA 1217 S1|CystA 1218 S1|CystA 1219 S1|CystA 1220 S1|CystA 1221 S1|CystA 1222 S1|CystA 1223 S1|CystA 1224 S1|CystA 1225 S1|CystA 1226 S1|CystA 1227 S1|CystA 1228 S1|CystA 1229 S1|CystA 1230 S1|CystA 1231 S1|CystA 1232 S1|CystA 1233 S1|CystA 1234 S1|CystA 1235 S1|CystA 1236 S1|CystA 1237 S1|CystA 1238 S1|CystA 1239 S1|CystA 1240 S1|CystA 1241 S1|CystA 1242 S1|CystA 1243 S1|CystA 1244 S1|CystA 1245 S1|CystA 1246 S1|CystA 1247 S1|CystA 1248 S1|CystA 1249 S1|CystA 1250 S1|CystA 1251 S1|CystA 1252 S1|CystA 1253 S1|CystA 1254 S1|CystA 1255 S1|CystA 1256 S1|CystA 1257 S1|CystA 1258 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