Methods for Plant RNA Isolation
时间:2005-07-18 00:00 来源:Scienceboard.net 作者:admin 点击:次
| Friday, November 07, 2003
Description Methods for Plant RNA Isolation Procedure AMES/Chloroform extraction in our lab to extract total RNA from potato leaves for viroid analysis. The protocol is as follows: AMES Buffer (for 200mL): 11.7g NaCl 160 mL dH2O Adjust pH to 6.0 0.40g MgCl2 8.21g Na-acetate 40mL EtOH 6.0g SDS We homogenize the plant material(usually leaves) and dilute the sap 1:5 in AMES We then incubate for 30min. @37oC followed by the addition of an equal vol.of chloroform.Vortex and store on ice. We then centrifuge the samples @ 3000rpm in a Sorvall ss-34 for 10min. Total RNA is contained in the upper aqueous layer. This protocol works well for us. We use it to extract total RNA for analysis with DIG-labelled cRNA probe for PSTVd (Potato Spindle Tuber Viroid). Recipes Supplies Tips (责任编辑：泉水) |