title: BioMark System with Dynamic Array 96.96 for miRNA RT-qPCR
status: Public on Feb 14 2018
contact.city: San Francisco
contact.first: Priscilla
contact.last: Hsue
contact.email: priscilla.hsue@ucsf.edu
contact.state: CA
contact.address: 1001 Potrero Ave, Room 5G4
contact.department: Medicine
contact.country: USA
contact.institute: University of California, San Francisco
contact.phone: 415-206-8257
organism: Homo sapiens
description: PCR assay
manufacture_protocol: Dynamic Arrays 96.96 were empty nano volume PCR chips. We used TaqMan miRNA Assays from Life Technologies and RNA samples converted to cDNA and then Preamplified with TaqMan miRNA Reverse Transcription Kit and MegaPlex Human RT Primer Pool Av2.1 and Pool Bv3.0. (Life Technologies, Foster City, CA) in a 384 well Thermal Cycler. The cDNA samples were PreAmplified using TaqMan PreAmp Master Mix and PreAmp Primers, Human Pool A v2.1 and Pool B v3.0 (Life Technologies, Foster City, CA). Preamplified cDNAs then loaded to Dynamic Array 96.96 after mixed with TaqMan Gene Expression Master Mix. TaqMan miRNA Assays were also loaded to Dynamic Array 96.96 chip and then qPCR analysis performed on BioMark System (Fluidigm).