1
Download Total RNA Isolation
Transcript
Total RNA Isolation Protocol-at-a-glance (Rev. 04) XS NucleoSpin® RNA XS 1 Supply sample 2 Lyse and homogenize cells Use up to 5 x 105 cultured cells or 5 mg tissue samples 100 µl RA1 2 µl TCEP Mix 3 Add Carrier RNA 5 µl Carrier RNA working solution Mix 4 Filtrate lysate (optional) 11,000 x g 30 s 5 Adjust RNA binding condition 6 Bind RNA 100 µl 70 % ethanol Mix Load lysate 11,000 x g 30 s 7 Desalt silica membrane 100 µl MDB 11,000 x g 30 s 8 Digest DNA 25 µl DNase reaction mixture RT 15 min 9 Wash and dry silica membrane 1st wash 100 µl RA2 RT, 2 min 11,000 x g, 30 s 2nd wash 400 µl RA3 11,000 x g, 2 min 3rd wash 200 µl RA3 11,000 x g, 2 min 10 Elute highly pure RNA 10 µl RNase-free H2O 11,000 x g 30 s 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] MN
