A method for DNA extraction from leaves in a 96 - well format   brew University of Jerusalem, The Faculty of Agriculture, P . O . Box 12, Rehovot Eyal Fridman, Tzili Pleban and Dani Zamir The He 76100, Israel . FAX: 972 - 8 - 9468265; Email: zamir@agri . huji . ac . il We olation describe here a microtiter - based method for rapid DNA tly as a genomic template for polymeras is that was developed for tom e chain reaction (PCR) . This method is useful for CR amplification and for it lo cts aimed at n (1, 2, 3), the 96 - well format described here can enhance day at reduced cost . The method was developed for mapping experiments in tomato but is o and disc is ing extended t 10 edure is channel pipet, a swinging - bucket a Add 100 m l of grinding buffer (using a multichannel pipet) . l roup successive plates before centrifugation) . - centrifuge – 4000 rpm for 15 min . - - - to primer specifications . - Add 3 m l loading dye to the wells and load 15 m l of the reaction on a 1 . 5% agarose gel . Using Owl D3 - 14 horizontal system (150 ml gel and four combs of 50 wells), it is possible to score the genotypes of 2 microtiter plates (192 samples) . Loading can be performed using the multichannel pipet and it is suggested to do so before the gel is carefully submerged in the electrophoresis buffer .   ato from leaf tissue that can be used direc large - scale genotyping performed in marker - assisted selection programs, the screening of transgenic plants by P fine mapping in large segregating populations . This latter application is necessary for high e resolution mapping of Mendelian genes or quantitative tra ci (QTL) in proj map based cloning . Although several protocols are available for small DNA extractio the productivity to hundreds of samples a also shown to be efficient for other crops, such as potat melon . A single leaf d from each individual plant us a single - hole paper punch that is placed in a 96 - obtaine flat - well microtiter plate . The leaf samples can be kept in a - 80 ° C freezer for periods and the final DNA product is sufficient for at leas PCRs . The entire proc performed in a 96 - well microtiter plate, using a multi entrifuge (Eppendorf; 5810) and a 96 - plate thermocycler (MJ PTC 225) . c Protocol   sing single - hole paper punch . Put each disc at the - Collect a single leaf disc per plant u bottom of a well (flat - bottom microtiter plate) .   - - Grind the leaves gently with a seed crusher (HyPure HSC - 200) using a rubber mallet (HyPure SCM - 100) . - Incubate at 60 ° C for 20 min . -