Targeted NGS Panel of Known or Suspected Cystic or Ciliopathy Genes
The Sequencing Platform involves utilizing a short-read targeted next-generation sequencing (tNGS) panel of up to 357 known and candidate genes for polycystic kidney/liver diseases and ciliopathies. The target enrichment technology and custom probe design utilized is Agilent SureSelect. Library preparation is performed utilizing NEBNext Ultra DNA Preparation Kit (E7370) to sheer via Covaris sonification. 500ng of Genomic DNA is prepared for each sample. The sheared fragments are ligated with NEBNext Dual Index Primers (E7600S) and purified per NEBNext Ultra DNA Preparation Protocol. The resulting libraies are QC'd with the Agilent Bioanalyzer, looking for a Final library size of ~200bp sized fragments. The resulting library is then pooled and captured utilizing Agilent SureSelect XT Probe Kit (G9611A) and hybridized incubated overnight. The resulting pooled libraries are purified, QC'ed via Agilent Bioanalyzer, and sent for sequencing.
NIDDK-phs004088
Emilie3_s_R2187