EVAg

Positive control for H5 and N8 real-time RT-PCR Positive controls for H5 and N8 real-time RT-PCR assays are in vitro transcribed RNA derived from strain A/Duck/England/36254/14(H5N8). The transcript contains the Open Reading Frame of the H5 gene (from nt 1000 to nt 1170) and N8 gene (from nt 960 to nt 1090) as negative standard. Each microtube contains 35.1011 copies of target sequences diluted in yeast tRNA and lyophilised. Reconstitution of transcribed RNA • Add 3500 µl of distilled water to obtain a solution at a concentration of 109 copies/µl. Store at -80°C. • Dilute in H2O to prepare a master bank at 2x106 copies/µl. Store at -80°C. • From this, prepare a working bank of reagent at 2x104 copies/µl in order to avoid freeze/thaw cycles. Working tubes may be stored at -20°C for less than one week.