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Apr
Hot Phenol Extraction Rna. Phenol alone retains 10-15 of water resulting in an equal loss of RNA. Spin down RNA 20 min. Grow yeast cells in desired medium to mid-exponential phase OD600 05-07. This comes from Currents Protocols in Molecular Biology and from Hahns web site.
Mini Hot Phenol Arabidopsis RNA extraction procedure ver. The second basic protocol allows rapid preparation of total cytoplasmic RNA by using a nonionic detergent to. Grind tissue 05 to 20g in liquid N2 and transfer to 30 ml tube. This comes from Currents Protocols in Molecular Biology and from Hahns web site. Dry pellet at room temp 20 min. It also is a good partner with phenol for denaturing proteins and it dissolves lipids.
Overdrying can lead to difficulty resuspending pellet At this point RNA can either be re-precipitated or purified further using Qiagen RNAeasy.
Yeast RNA Extraction Using Hot Phenol See Köhrer and Domdey 1991 Methods Enzymol 194398-405. Because this procedure does not require vortexing individual samples with. At 15-20000g 12500 rpm in SS34 rotor. Prepare dry ice powder in bucket. Total RNA Extraction from Saccharomyces cerevisiae Using Hot Acid Phenol Molecular Cloning also known as Maniatis has served as the foundation of technical expertise in labs worldwide for 30 years. The separation of the organic phase from the aqueous phase is achieved using Phase Lock Gel an inert material with a density intermediate between the organic and aqueous samples.