Gene Expression:Article Title: Synergy between HA cleavage site sequence and NA-mediated plasminogen recruitment as a virulence mechanism for low-pathogenic avian influenza.
Article Snippet: .. Analysis of gene expression via 96.96 IFC Fluidigm Dynamic Array qPCR cDNA was synthesized from 200 ng of RNA using a high-capacity reverse transcrip tion kit (Applied Biosystems) according to the manufacturer’s instructions. cDNA was Month XXXX Volume 0 Issue 0 10.1128/mbio.02466-25 6 D ow nl oa de d fr om h ttp s: //j ou rn al s. as m .o rg /jo ur na l/m bi o on 0 2 M ar ch 2 02 6 by 1 50 .1 65 .2 54 .2 7. pre-amplified with TaqMan Pre-Amp Master Mix (Applied Biosystems) and 2.5 μL of a 200 nM mixed pool of primer pairs (as described previously [34]; see Table S1 for the gene list used here) at 95°C for 10 min, followed by 14 cycles of 95°C for 15 s and 60°C for 4 min. Pre-amplified cDNA was treated with 16 U/μL Exonuclease I (New England Biolabs) at 37°C for 30 min before heat inactivation at 80°C for 15 min. Pre-amplified, exonucleasetreated cDNA was analyzed through qPCR with the microfluidic 96.96 Dynamic Fluidigm array on the BioMark HD instrument (BioMark). .. Assay mixes were prepared with 2.25 μL 2× assay loading reagent (Fluidigm), 2.5 μL of primer pair mix (1.15 μM), and 0.25 μL low EDTA TE buffer.
Real-time Polymerase Chain Reaction:Article Title: Synergy between HA cleavage site sequence and NA-mediated plasminogen recruitment as a virulence mechanism for low-pathogenic avian influenza.
Article Snippet: .. Analysis of gene expression via 96.96 IFC Fluidigm Dynamic Array qPCR cDNA was synthesized from 200 ng of RNA using a high-capacity reverse transcrip tion kit (Applied Biosystems) according to the manufacturer’s instructions. cDNA was Month XXXX Volume 0 Issue 0 10.1128/mbio.02466-25 6 D ow nl oa de d fr om h ttp s: //j ou rn al s. as m .o rg /jo ur na l/m bi o on 0 2 M ar ch 2 02 6 by 1 50 .1 65 .2 54 .2 7. pre-amplified with TaqMan Pre-Amp Master Mix (Applied Biosystems) and 2.5 μL of a 200 nM mixed pool of primer pairs (as described previously [34]; see Table S1 for the gene list used here) at 95°C for 10 min, followed by 14 cycles of 95°C for 15 s and 60°C for 4 min. Pre-amplified cDNA was treated with 16 U/μL Exonuclease I (New England Biolabs) at 37°C for 30 min before heat inactivation at 80°C for 15 min. Pre-amplified, exonucleasetreated cDNA was analyzed through qPCR with the microfluidic 96.96 Dynamic Fluidigm array on the BioMark HD instrument (BioMark). .. Assay mixes were prepared with 2.25 μL 2× assay loading reagent (Fluidigm), 2.5 μL of primer pair mix (1.15 μM), and 0.25 μL low EDTA TE buffer.
Synthesized:Article Title: Synergy between HA cleavage site sequence and NA-mediated plasminogen recruitment as a virulence mechanism for low-pathogenic avian influenza.
Article Snippet: .. Analysis of gene expression via 96.96 IFC Fluidigm Dynamic Array qPCR cDNA was synthesized from 200 ng of RNA using a high-capacity reverse transcrip tion kit (Applied Biosystems) according to the manufacturer’s instructions. cDNA was Month XXXX Volume 0 Issue 0 10.1128/mbio.02466-25 6 D ow nl oa de d fr om h ttp s: //j ou rn al s. as m .o rg /jo ur na l/m bi o on 0 2 M ar ch 2 02 6 by 1 50 .1 65 .2 54 .2 7. pre-amplified with TaqMan Pre-Amp Master Mix (Applied Biosystems) and 2.5 μL of a 200 nM mixed pool of primer pairs (as described previously [34]; see Table S1 for the gene list used here) at 95°C for 10 min, followed by 14 cycles of 95°C for 15 s and 60°C for 4 min. Pre-amplified cDNA was treated with 16 U/μL Exonuclease I (New England Biolabs) at 37°C for 30 min before heat inactivation at 80°C for 15 min. Pre-amplified, exonucleasetreated cDNA was analyzed through qPCR with the microfluidic 96.96 Dynamic Fluidigm array on the BioMark HD instrument (BioMark). .. Assay mixes were prepared with 2.25 μL 2× assay loading reagent (Fluidigm), 2.5 μL of primer pair mix (1.15 μM), and 0.25 μL low EDTA TE buffer.
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