Materials and Reagents

1. Agencourt Chloropure Kit (this kit is discontinued by Beckman Coulter but can be ordered) or Quiagen MagAttract 96 DNA plant core kit (QIAGEN, catalog number: 67163 )
   
2. Qiagen Type-it Multiplex PCR kit (QIAGEN, catalog number: 206243 )
   
3. QIAquick PCR purification kit (QIAGEN, catalog number: 28106 )
   
4. Liquid nitrogen
   
5. 100% Isopropanol, ultrapure
   
6. Freshly prepared 70% ethanol made with nuclease free water
   Note: 70% ethanol is hygroscopic, always make fresh for optimal result.
   
7. Agarose
   
8. Ethydium bromide solution
   
9. Nuclease free water
   
10. Pure molecular biology grade ethanol (96– 100%)
    Note: Calculate the volume of ethanol needed before you start. A new kit of 24 plates needs about 5 L of ethanol.
    
11. GENSCAN 500 ROX (Applied Biosystems, catalog number: 401734 )
12. 1x TAE (Tris-Acetate-EDTA) buffer (see Recipes)
    

Equipment

1. 2.2 ml Ritter Deep-well plates (ABGene, catalog number: AB-6661 )
   
2. Adhesive plate film (ABGene, catalog number: AB-0558 , AB-626 , AB-662 )
   
3. 4 mm steel balls (SPEX SamplePrep, catalog number: 2150 )
   
4. Pipette tips
   
5. Processsing Plate: Deep-well titer plate with a 96-well format (SPEX SamplePrep, catalog number: 2210 )
   
6. Destination Plate: 300 μl round bottom microtiter plate (96 well, 300 μl well capacity, round bottom) (Corning, Costar^®, catalog number: 07-200-105 )
   
7. Desktop centrifuge for 96 well plates
   
8. Reagent reservoir
   
9. Genogrinder (SPEX SamplePrep, catalog number: 2010 )
   
10. Cooling block (SPEX SamplePrep, catalog number: 2665 )
    
11. Steel ball dispenser (SPEX SamplePrep, catalog number: 2100 )
    
12. Multi-channel pipettes (0.1-10, 10-100, 30-300 μl)
    
13. Agencourt Supermagnet magnetic Plate (Agencourt SPRIPlate 96R-Ring Magnet Plate, catalog number: A29164 )
    
14. BioRad Gel casting tray, running tray, power pack etc (Bio-Rad Laboratories)
    
15. Nanodrop spectrophotometer (Nanodrop)
    
16. Pipette (0.2-2 μl), pipette tips, soft tissue paper
    
17. ABI 3100 Genetic Analyzer (Applied Biosysyems)
    

Procedure