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Download Total RNA Isolation
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Total RNA Isolation Protocol-at-a-glance (Rev. 11) Mini Midi NucleoSpin RNA II ® 1 Homogenize sample 2 Lyse cells 3 NucleoSpin® RNA L 30 mg 100 mg 350 µl RA1 3.5 µl ß-mercaptoethanol 1.8 ml RA1 18 µl ß-mercaptoethanol Mix Mix Filtrate lysate 11,000 x g 1 min 4 5 6 7 8 Adjust RNA binding conditions 1.8 ml 70 % ethanol Mix Mix Desalt silica membrane Wash and dry silica membrane Elute highly pure RNA Load sample Load sample 11,000 x g 30 s 4,500 x g 3 min 350 µl MDB 2.2 ml MDB 11,000 x g 1 min 4,500 x g 3 min 95 µl DNase reaction mixture 250 µl DNase reaction mixture RT, 15 min RT, 15 min 1st wash 200 µl RA2 1st wash 2.6 ml RA2 2nd wash 600 µl RA3 2nd wash 2.6 ml RA3 3 wash 250 µl RA3 3 wash 2.6 ml RA3 rd 9 350 µl 70 % ethanol Bind RNA Digest DNA 4,500 x g 10 min rd 1st and 2nd 11,000 x g 30 s 1st and 2nd 4,500 x g 3 min 3rd 11,000 x g 2 min 3rd 4,500 x g 5 min 60 µl RNasefree H2O 11,000 x g 1 min MACHEREY-NAGEL GmbH & Co. KG • Neumann-Neander-Str. 6-8 • D-52355 Düren • Germany Tel.: +49 (0) 24 21 969 270 • www.mn-net.com • e-mail: [email protected] 500 µl RNasefree H2O RT, 2 min 4,500 x g 3 min MN
