植物RNA提取--Methods for Plant RNA Isolation

 AMES/Chloroform extraction in our lab to extract total 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. @37℃ 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 is contained in the upper aqueous layer. This protocol works well for us. We use it to extract total for analysis with DIG-labelled c probe for PSTVd (Potato Spindle Tuber Viroid).