Materials to Be Supplied by the User (Solution compositions are provided in Section 8.A.) 100% isopropanol, RNase-free 95% ethanol, RNase-free microcentrifuge 4.A. Preparation of Solutions Before beginning the ReliaPrep RNA Tissue Miniprep System protocol, four solutions must be prepared.
Miniprep Protocol . Points of Emphasis: The entire miniprep procedure takes 40-45 min. The video shows purification of cDNA from one bacterial culture. Normally, 9-18 cultures are processed at one time so the procedure will take longer, especially for beginners. Use proper pipetting techniques to insure that that 1000 µl pipet does not become
Bioone.org Overview of protocols —Option 1 is the manufacturer's protocol for RNA extraction with TRIzol (Ambion, Life Technologies, Carlsbad, California, USA), with the addition of sarkosyl to the extraction solution. Option 2 combines traditional cetyltrimethylammonium bromide (CTAB) extraction (Doyle and Doyle, 1987) followed by the use of ...
USA), 20ng/mL EGF (Invitrogen, Carlsbad, CA, USA), and 20ng/mL FGF (Invitrogen, Carlsbad, CA, USA). Real-time quantitative reverse transcriptase PCR Total RNA was extracted using the Direct-zol™ RNA MiniPrep kit (Zymo Research, CA, USA) from TRIsure RNA extraction reagent. The synthesis of complementary