other:
RNA Extraction:Article Title: Use of inhaled ciclesonide for treatment of moderate asthma in Thoroughbred racehorses.
Article Snippet: .. All Tempus blood RNA tubes were processed for total RNA extraction at the Gluck Equine Research Center, in Lexington, Kentucky as previously described.29 Briefly, RNA was isolated using a KingFisher Flex (Thermo Fisher Scientific Inc, Massachusetts, USA) and the MagMax Core kit (Applied Biosystems, California, USA) per the manufacturer's recommendations, except there was no DNase step and the pelleted RNA was resuspended in 600 μL viral lysis buffer (Invitrogen Corporation, California, USA). .. Expression of the following genes was calculated using β-glucuronidase (β-Gus), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyltransferase 1 (HPRT1) and Intercellular Adhesion Molecule 1 (ICAM1) as reference genes for all the samples using the relative expression software tool (REST) as described.12,30 The use of REST allowed for correction for PCR efficiency and normalization using multiple reference genes.12 Housekeeping genes were selected based on the stability of their expression.31 Commercially available primers and probes were used (Thermo Fisher Scientific): β glucuronidase (β-GUS, Ec03470630_m1), glyceraldehyde 3-phosphate dehydrogenase (GAPDH, Ec03210916_gH), hypoxanthine phosphoribosyltransferase 1 (HPRT1, Ec03470217_m1), Intercellular Adhesion Molecule 1 (ICAM1, Ec07080735_m1), Interferon-γ (IFNγ, Ec03468606_m1), Interleukins (IL), IL-1β (Ec04260298_s1), IL-4 (Ec03468864_m1), IL-6 (Ec03468678_m1), IL-8 (Ec03468860_m1), IL-10 (Ec03468647_m1), IL-12A (Ec03468747_m1), IL-12B (Ec03468777_m1), IL-13 (Ec03470543_m1), IL-17A (Ec03470096_m1), IL-23A (Ec03468800_m1), and tumor necrosis factor-α (TNFα, Ec03467871_m1).
Isolation:Article Title: Use of inhaled ciclesonide for treatment of moderate asthma in Thoroughbred racehorses.
Article Snippet: .. All Tempus blood RNA tubes were processed for total RNA extraction at the Gluck Equine Research Center, in Lexington, Kentucky as previously described.29 Briefly, RNA was isolated using a KingFisher Flex (Thermo Fisher Scientific Inc, Massachusetts, USA) and the MagMax Core kit (Applied Biosystems, California, USA) per the manufacturer's recommendations, except there was no DNase step and the pelleted RNA was resuspended in 600 μL viral lysis buffer (Invitrogen Corporation, California, USA). .. Expression of the following genes was calculated using β-glucuronidase (β-Gus), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyltransferase 1 (HPRT1) and Intercellular Adhesion Molecule 1 (ICAM1) as reference genes for all the samples using the relative expression software tool (REST) as described.12,30 The use of REST allowed for correction for PCR efficiency and normalization using multiple reference genes.12 Housekeeping genes were selected based on the stability of their expression.31 Commercially available primers and probes were used (Thermo Fisher Scientific): β glucuronidase (β-GUS, Ec03470630_m1), glyceraldehyde 3-phosphate dehydrogenase (GAPDH, Ec03210916_gH), hypoxanthine phosphoribosyltransferase 1 (HPRT1, Ec03470217_m1), Intercellular Adhesion Molecule 1 (ICAM1, Ec07080735_m1), Interferon-γ (IFNγ, Ec03468606_m1), Interleukins (IL), IL-1β (Ec04260298_s1), IL-4 (Ec03468864_m1), IL-6 (Ec03468678_m1), IL-8 (Ec03468860_m1), IL-10 (Ec03468647_m1), IL-12A (Ec03468747_m1), IL-12B (Ec03468777_m1), IL-13 (Ec03470543_m1), IL-17A (Ec03470096_m1), IL-23A (Ec03468800_m1), and tumor necrosis factor-α (TNFα, Ec03467871_m1).
Lysis:Article Title: Use of inhaled ciclesonide for treatment of moderate asthma in Thoroughbred racehorses.
Article Snippet: .. All Tempus blood RNA tubes were processed for total RNA extraction at the Gluck Equine Research Center, in Lexington, Kentucky as previously described.29 Briefly, RNA was isolated using a KingFisher Flex (Thermo Fisher Scientific Inc, Massachusetts, USA) and the MagMax Core kit (Applied Biosystems, California, USA) per the manufacturer's recommendations, except there was no DNase step and the pelleted RNA was resuspended in 600 μL viral lysis buffer (Invitrogen Corporation, California, USA). .. Expression of the following genes was calculated using β-glucuronidase (β-Gus), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine phosphoribosyltransferase 1 (HPRT1) and Intercellular Adhesion Molecule 1 (ICAM1) as reference genes for all the samples using the relative expression software tool (REST) as described.12,30 The use of REST allowed for correction for PCR efficiency and normalization using multiple reference genes.12 Housekeeping genes were selected based on the stability of their expression.31 Commercially available primers and probes were used (Thermo Fisher Scientific): β glucuronidase (β-GUS, Ec03470630_m1), glyceraldehyde 3-phosphate dehydrogenase (GAPDH, Ec03210916_gH), hypoxanthine phosphoribosyltransferase 1 (HPRT1, Ec03470217_m1), Intercellular Adhesion Molecule 1 (ICAM1, Ec07080735_m1), Interferon-γ (IFNγ, Ec03468606_m1), Interleukins (IL), IL-1β (Ec04260298_s1), IL-4 (Ec03468864_m1), IL-6 (Ec03468678_m1), IL-8 (Ec03468860_m1), IL-10 (Ec03468647_m1), IL-12A (Ec03468747_m1), IL-12B (Ec03468777_m1), IL-13 (Ec03470543_m1), IL-17A (Ec03470096_m1), IL-23A (Ec03468800_m1), and tumor necrosis factor-α (TNFα, Ec03467871_m1).
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