Universal probe library algorithm

RNA was purified from whole cell lysates using the RNeasy Mini kit (Qiagen) or the Illustra Triple Prep kit (GE Healthcare). RNA was reverse transcribed using High Capacity cDNA Reverse Transcription kit (Applied Biosystems) with random hexamer priming. Gene-specific primers were used in combination with probes designed using the Universal Probe Library algorithm (Roche) and Universal Master mix (Roche); all reactions were run on a Mastercycler Realplex2 system (Eppendorf). Relative quantification to the control (cyclophilin B) was done using the comparative ΔΔCT method. Individual sample PCR reactions were performed in triplicate. The following gene specific primers (Integrated DNA Technologies) were used: Cyclophilin B Forward – 5’-ttcttcataaccacagtcaagacc-3’, Cyclophilin B Reverse – 5’-accttccgtaccacatccat-3’, UPL 20; MV nucleoprotein Forward - 5’-ggaaactgcaccctacatgg-3’, MV nucleoprotein Reverse- 5’-gggtatgatcctgcactgaact-3’, UPL 80; Bst2 Forward- 5’-gaagtcacgaagctgaacca-3’, Bst2 Reverse- 5’-cctgcactgtgctagaagtctc-3’, UPL 78.

RNA was purified from homogenized whole brain tissue (see below) and quantified using a Nanodrop spectrophotometer. RNA was reverse-transcribed using High Capacity cDNA Reverse Transcription kit (Applied Biosystems) with random hexamer priming. Gene-specific primers were used in combination with probes designed using the Universal Probe Library algorithm (Roche) and Universal Master mix (Roche); all reactions were run on a Mastercycler Realplex2 system (Eppendorf). Cycling conditions were 50°C, 2 min; 95°C, 10 min; followed by 40 (2-step) cycles (95°C, 15 sec; 60°C, 60 sec). Relative quantification to the control (cyclophilin B) was done using the comparative ΔΔCT method. Individual sample PCR reactions were performed in triplicate. The following gene specific primers (Integrated DNA Technologies) were used: Cyclophilin B Forward – 5′-ttcttcataaccacagtcaagacc-3′, Cyclophilin B Reverse – 5′-accttccgtaccacatccat-3′, UPL 20; MV nucleoprotein Forward - 5′-ggaaactgcaccctacatgg-3′, MV nucleoprotein Reverse- 5′-gggtatgatcctgcactgaact-3′, UPL 80.