, Transcribe 1 µg of linearized DNA using an mRNA synthesis kit with the appropriate polymerase, according to manufacturer's instructions

, 7):chloroform and ammonium acetate stop solution provided with the kit, Extract the mRNA with, vol.5, p.1

, Vortex for 20 sec, centrifuge 10 min at 18,000 x g and collect the aqueous phase

, Extract the aqueous phase again with 24:1 chloroform:isoamyl alcohol. Vortex for 20 sec, centrifuge 10 min at 18,000 x g and collect the aqueous phase

, Precipitate the mRNA with 100% isopropanol overnight at-20 °C

, Let the pellet dry at room temperature for no longer than 5-10 min as it will then be difficult to resuspend. Resuspend in DNase-and RNase-free water to a final concentration of at least 2 µg/µl to ensure a decent final mRNA concentration in the injection mix, Centrifuge 10 min at 18,000 x g at 4 °C

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