GSM5589589: Normal-Cancer-freeP57; Homo sapiens; miRNA-Seq
Blood samples from NHD and cancer patients were obtained. All blood samples were collected in Streck tubes (Streck, La Vista, NE, USA) or EDTA tubes (BD vacutainers®, USA). Blood samples were processed and cryopreserved at -80°C. All plasma samples were analyzed by the HTG miR WTA. Libraries were prepared according to Illumina instrument sequencing protocols. Probe-captured miR samples were amplified and indexed via polymerase chain reaction (PCR) using master mix (dNTP 10 mM, 5X Hemo Klentaq® Reaction buffer, Hemo Klentaq® DNA polymerase (New England Biolabs Inc., Ipswich, MA), forward, and reverse primers (HTG Molecular Diagnostics Inc., Tucson, AZ).  Following PCR, library cleanup was performed using Ampure® XP beads (Beckman Coulter Inc., Brea, CA, USA). The samples were mixed  and incubated with the beads at room temperature  for 5 min. The bead-bound libraries were then  washed twice with 80% ethanol. After washing, the samples were dried at RT for 2 min. The libraries were then  eluted off the beads with 10 mM Tris hydrochloride, pH 8.0.