METHOD FOR SINGLE CELL ISOLATION AND RNA EXTRACTION FROM WOODY TISSUES

摘要

The present patent application discloses a method for single cell isolation and RNA extraction in woody plants' tissues, while preserving the viability of the nucleic acids for further analysis. The method comprises, collecting a woody plant tissue sample, snap-freezing and embedding the sample in Optimal Cutting Temperature (OCT) freezing medium, sectioning of the frozen samples into sections of 10 to 20 µm of thickness, fixation and dehydration of plant sections, isolation of single cells by laser microdissection, RNA extraction from isolated single-cells, wherein the RNA extraction comprises the use of a lysis buffer comprising stabilizers.;Thus, it is disclosed here for the first time a method for single cell isolation and RNA extraction of isolated woody tissues comprising xylem and suberized cells while still maintaining RNA integrity for RNA sequence analysis.