A simple protocol to purify fresh nuclei from milligram amounts of meristematic pea root tissue for biochemical and flow cytometry applications

Abstract

A simple protocol to purify fresh nuclei from very small amounts (mg) of Pisum sativum (cv. Lincoln) root tissue is presented. The protocol is reliable and has been repeated many times; it can be performed within a few hours and needs only a simple modification of common glassware for tissue homogenization. Similar yields of purified nuclei are obtained both with meristematic and adult root tissues using a detergent‐free glyeerol buffer. Purified nuclei can be stored at –20°C for several weeks without any appreciable loss of integrity. The high yield of purified nuclei enables us to use our preparations for biochemical and cytometric investigations.