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Table 1 PCR conditions for quantitative real-time PCR analysis of hamster and mouse models of TSE disease.

From: Differential expression of interferon responsive genes in rodent models of transmissible spongiform encephalopathy disease

Hamster
Gene Primer Pairs Size (bp) Denature 95°C (sec) Anneal (°C) Anneal Time (sec) Extension 72°C (sec) Mg2+ (mM) Number Cycles
Mx2 5'-TGGCGGTAGGCATTCAGG-3' 5'-TGCCAGGACCAAGTTTACAGG-3' 161 5 54 5 7 4 40
2'5'-OAS 5'-AGCCGCTGCCCCCACTGTA-3' 5'-TCCGAGACTGCCCTGAAGC-3' 109 5 57 5 5 3 50
Eif2ak2 5'-TAGGCCTTGTCAACAGTTATGCTCA-3' 5'-GCTGCTTTGCCTCCTGCTTGGTAG-3' 162 5 58 5 7 3 45
GAPdH 5'-ATGGCAAGTTCAAAGGCACAGTCA-3' 5'-TGGGGGCATCAGCAGAAGG-3' 231 5 60 5 11 3 40
GFAP 5'-GGACATCGAGATTGCCACCTAT-3' 5'-CATCCCTCATCTCCACTGTCTTTA-3' 176 5 60 5 8 3 45
Mouse
Gene Primer Pairs Size (bp) Denature 95°C (sec) Anneal (°C) Anneal Time (sec) Extension 72°C (sec) Mg2+ (mM) Number Cycles
Mx2 5'-CCTGCCTGCCATCGCTGTC-3' 5'-GCCTCTCCACTCCTCTCCCTCATT-3' 160 5 60 5 11 3 40
2'5'-OAS 5'-GAGGCGGTTGGCTGAAGAGG-3' 5'-GAGGAAGGCTGGCTGTGATTGG-3' 312 5 60 5 11 3 40
Eif2ak2 5'-GTACAAGCGCTGGCAGAACTCAAT-3' 5'-AAGAGGCACCGGGTTTTGTAT-3' 124 5 60 5 11 4 40
GAPdH 5'-CACGGCAAATTCAACGGCACAGT-3' 5'-TGGGGGCATCGGCAGAAGG-3' 232 5 60 5 11 3 40
GFAP 5'-GAGCGAGCGTGCAGAGATGATGG-3' 5'-CTCCCGAAGCTCCGCCTGGTAGA-3' 166 5 60 5 11 3 45
Caspase 8 5'-TCTGCTGGGAATGGCTACGGTGAA-3' 5'-GTGTGAAGGTGGGCTGTGGCATCT-3' 212 5 60 5 11 3 45
TNFα 5'-TGCTCTGTGAAGGGAATGGGTGTT-3' 5'-AGTCCTTGATGGTGGTGCATGAGA-3' 241 5 60 5 11 3 45
TNFα Receptor 5'-AACCAGTTCCAACGCTACCTGAGT-3' 5'-AAGGGACGCACTCACTTTCTCTCA-3' 157 5 60 5 8 3 45
Fas Ligand 5'-ATCCCTCTGGAATGGGAAGACACA-3' 5'-ACCCAGTTTCGTTGATCACAAGGC-3' 91 5 63 5 6 3 45
Fas/ApoI Receptor 5'-TCGCCTATGGTTGTTGACCATCCT-3' 5'-TGGTATGGTTTCACGACTGGAGGT-3' 136 5 57 5 7 3 45
  1. Conditions outlined above were used to determine the relative expression levels of the genes Eif2ak2, 2'5'-OAS, Mx2, GFAP, TNFα, TNFR1, FasL, Fas/ApoI, and caspase 8, as well as the reference gene GAPdH using the Roche LightCycler® 1.5.