Pooled sample consisting of samples: Cyst 541 S1|Cyst 558 S1|Cyst 590 S1|Cyst 503 S1|Cyst 543 S1|Cyst 559 S1|Cyst 567 S1|Cyst 575 S1|Cyst 591 S1|Cyst 585 S1|Cyst 530 S1|Cyst 538 S1|Cyst 554 S1|Cyst 515 S1|Cyst 531 S1|Cyst 563 S1|Cyst 587 S1|Cyst 516 S1|Cyst 548 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