肥厚性梗阻型心肌病伴持续性st段抬高一例.docVIP

JTRM-D-17-00060 Novel Trigenic CACNA1C/DES/MYPN Mutations in a Family of Hypertrophic Cardiomyopathy with Early Repolarization and Short QT Syndrome Short Title – Chen, et al: Novel Trigenic Mutations in HCM family ER and SQTS Online Supplement Online Methods Clinical history The study was approved by the ethics committee of 3rd People’s Hospital of Wuxi (Wuxi, China) and conducted according to Declaration of Helsinki principles. The informed consents were obtained from all the participants, who belonged to Asian. The clinical assessments included history collection, detailed physical examination, blood tests, electrocardiogram (ECG), ultrasonic cardiogram (UCG), and coronary artery computer tomography angiography. Patients were clinically diagnosed according to the 2011 ACC/AHA Guideline for the Diagnosis and Treatment of Hypertrophic Cardiomyopathy[1]. Genetic studies DNA extraction, Target region capture, and Next Generation Sequencing Genomic DNA was extracted from peripheral blood lymphocytes by standard procedures using QIAamp DNA Bloodmini kits (Qiagen, Germany). One μg genomic DNA was fragmented by Covarissonicator (Covaris S2, USA) to sizes of 150-300 bp and then purified. The blunt ends of the purified DNA fragments were repaired, and A-tailing was added. The fragments were ligated overnight using standard Illumina paired- end (PE) adapter. The ligated products were then amplified through 4-cycle polymerase chain reactions (PCRs) using PE primers containing 8 bp index tags. The purified PCR products containing 3 μg DNA were hybridized to the GenCapTM probe (in solution) at 65°C for 22 hours using a PCR machine. The products were bound to a rotator for 1 hour at room temperature using DynalMyone Streptavidin C1 magnetic beads (Invitrogen, USA), which had been activated beforehand, and the products were then washed with buffer according to the kit manual. The captured DNA libraries were amplified using 15-cycle PCRs, purified, and subsequently eluted in a 3