title: nCounter Custom CodeSet
status: Public on Oct 10 2019
contact.city: Auckland
contact.first: Tina
contact.last: Nie
contact.email: t.nie@auckland.ac.nz
contact.address: 3 Symonds St
contact.department: School of Biological Sciences
contact.country: New Zealand
contact.institute: The University of Auckland
organism: Mus musculus
manufacture_protocol: A hybridisation master mix was made by adding 70 µL of hybridisation buffer to the Reporter CodeSet provided. 8 µL of this was added to a strip tube. To each tube, 5 µL of RNA sample was added. Note the manufacturer’s protocol calls for 100 ng of total RNA. We added 5 µL of our sample without dilution (giving a greater chance of counting low expression genes). Variable amounts of RNA were normalised to the chosen reference genes. 2 µL of the Capture Probeset was pipetted into each tube. The tubes were spun down briefly before being placed in thermocycler set at 65° C. Samples were left to incubate for 17 hours. The reaction was stopped by cooling samples to 4° C. Samples were added to a slide and run on the GEN2 Prep Station. The high sensitivity setting was used in our experiment. Slides were imaged on the Digital Analyser using 280 field-of-view counts.