184.108.40.206. RNA and cDNA preparation
- Centrifuge thawed aliquots for 2 min at 6,500 x g and 4°C.
- Resuspend bacterial pellets in TE buffer with lysozyme (15 mg/ml) and proteinase K (Qiagen).
- Vortex for 10 sec.
- Incubate 10 min at RT with constant shaking.
- Isolate RNA with RNeasy Plus Mini Kit and protocol “Purification of Total RNA for Animal Cells” (with on-column DNase I treatment).
- Elute RNA with 30 µl RNase-free water.
- Store at -80°C.
- Convert RNA to cDNA with RevertAid First Strand cDNA Synthesis Kit (Fermentas), using random hexamer primers.