Selected article for: "free water and RT reaction"
Author: Barker, Emily N.; Stranieri, Angelica; Helps, Chris R.; Porter, Emily L.; Davidson, Andrew D.; Day, Michael J.; Knowles, Toby; Kipar, Anja; Tasker, Séverine
Title: Limitations of using feline coronavirus spike protein gene mutations to diagnose feline infectious peritonitis
  • Document date: 2017_10_5
    • ID: 08b0g46x_15
    Snippet: Reverse transcription was performed using a MJ Mini Gradient Thermal Cycler and ImProm II Reverse Transcriptase (Promega). Ten microlitre of total RNA were combined with ImProm II 5 × Reaction Buffer, 3 mM MgCl 2 , dNTPs (0.5 mM each), random hexamers (25 ng/ μL) and ImProm II reverse transcriptase in a total volume of 20 μL. The following thermal profile was used; 20 °C for 5 min, 42 °C for 30 min, 70 °C for 15 min and 10 °C hold. The res.....
    Document: Reverse transcription was performed using a MJ Mini Gradient Thermal Cycler and ImProm II Reverse Transcriptase (Promega). Ten microlitre of total RNA were combined with ImProm II 5 × Reaction Buffer, 3 mM MgCl 2 , dNTPs (0.5 mM each), random hexamers (25 ng/ μL) and ImProm II reverse transcriptase in a total volume of 20 μL. The following thermal profile was used; 20 °C for 5 min, 42 °C for 30 min, 70 °C for 15 min and 10 °C hold. The resulting 20 μL of cDNA was added to 30 μL of RNase-free water and stored at −20 °C. Randomly selected samples were checked for inhibition of the RT reaction using an RNA internal amplification control. No inhibition was detected (results not shown).

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