RNA fraction WASH 3X 0.3M guanidine hydrochloride in 95% EtOH

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RNA fraction WASH 3X 0.3M guanidine hydrochloride in 95% EtOH 1 X 100% EtOH DNA fraction +Trizol® +Chloroform homogenization 12.000 g 4⁰C, 15 min incubation at RT 5 min +Ethanol 7.500 g 4⁰C, 5 min 2.000 g 4⁰C, 15 min + 8M urea/ 2M thiourea (UT) Thawed sample +Isopropanol 12.000 g 4⁰C, 15 min DNA precipitation Discard supernatant 10.000 g 4⁰C, 10 min Protein precipitation Protein pellet Incubate 20⁰C Shake 800 rpm Collect supernatant Protein extract solution Supplementary Fig. S1 The Trizol protocol according to the manufacturer’s protocol with a modified protein extraction work flow. The modified steps focus on an improved reconstitution of the protein pellet in 8M urea/2M thiourea (UT) and are highlighted by blue background.

+Trizol® homogenization incubation at RT 5 min +Chloroform RNA +Ethanol +Isopropanol Thawed sample 12.000 g 4⁰C, 15 min DNA precipitation Discard pellet 2.000 g Protein precipitation Discard supernatant WASH 3X 0.3M guanidine hydrochloride in 95% EtOH 1 X EtOH 100% 7.500 g 4⁰C, 5 min +150µl 8M urea/ 2M thiourea (UT) Incubate 20⁰C 800 rpm Protein pellet 10.000 g 4⁰C, 10 min Protein extract solution Collect supernatant Figure 1. The modified protein extraction protocol with Trizol, also depicting the dissolution step by addition of ~150µl 8M urea/2M thiourea (UT) to each sample