Microprep protocol for extraction of DNA from tomato and other herbaceous plants

Abstract

T he extraction of DNA from plant tissue is a critical and often very time-consuming step in many plant molecular biology procedures. This is especially true for studies of molecular genetics, QTLs, or molecular-marker-based breeding where hundreds or even thousands of plant samples need to be analyzed in a short period of time. Many protocols are laborious and are limited by the need for large amounts of plant tissue. Based on the methods originally described by Murray et al. (1980), we have developed a procedure in our laboratory that maximizes the number of plant samples one person can extract and that yields sufficient DNA for 50 to 100 PCR reactions or two to four Southern blots. The use of very small, new leaves makes it possible to extract DNA from seedlings only one to three weeks old, reducing the need for large amounts of greenhouse space. The entire procedure can be done in 1.5-mL microcentrifuge tubes, eliminating the need for large centrifuges. Using this procedure, one person can isolate DNA from several hundred plants per day. Materials and Solut ions