Maize Cell Disruption Methods


	Maize Cell Disruption Methods

Home Products Online Store How to Order International Distributors Contract Services About Us Contact Us Links Sales Opportunities OPS Diagnostics, LLC 291 Route 22 East Salem Industrial Park Building 6 Lebanon, NJ 08833 TEL: (908) 253-3444 FAX: (908) 575-1660 Email: info@opsdiagnostics.com Distribution Opportunities		Home > Application Notes >Application Table Maize can be pulverized by employing several methods depending on the part of the plant and the desired end result. Adult leaves, dry kernels, and stems are some of the toughest parts to homogenize into fine powders, because of their durability and toughness. Yet young seedlings may be less challenging and can be done in a 96 well format. They may require a combination of techniques including; treatment with liquid nitrogen, pulverizing by bead beating, or freeze drying and grinding with a mortar and pestle to render the best overall outcome. References McMullen, Michael D., Stephen Kresovich, Hector Sanchez Villeda, Peter Bradbury, Huihui Li, Qi Sun, Sherry Flint-Garcia, Jeffry Thornsberry, Charlotte Acharya, Christopher Bottoms, Patrick Brown, Chris Browne, Magen Eller, Kate Guill, Carlos Harjes, Dallas Kroon, Nick Lepak, Sharon E. Mitchell, Brooke Peterson, Gael Pressoir, Susan Romero, Marco Oropeza Rosas, Stella Salvo, Heather Yates, Mark Hanson, Elizabeth Jones, Stephen Smith, Jeffrey C. Glaubitz, Major Goodman, Doreen Ware, James B. Holland, Edward S. Buckler. 2009. Genetic Properties of the Maize Nested Association Mapping Population. Science 325, 737 Protocols Single and Multiple Dry and Wet Kernel Bead Beating Method Place one dry kernel into a 4 ml vial. Depending on the durometer of the kernel, either a 3/8" stainless steel or tungsten carbide ball can be added to the vial. Cap the vial, place it into the Geno/Grinder or HT Homogenizer and homogenize for 2 minutes on high. Samples consisting of up to fifteen kernels can be homogenized in 15 ml vials with two 7/16" stainless steel or tungsten balls for similar results as the single kernel 4ml format. Wet or hydrated kernels should be homogenized with 100ul of buffer solution added to the tube. *Other Considerations* Bead beating generates heat which may break down certain enzymes. Tungsten carbide is best for severely dried kernels, because it is almost twice the density of stainless steel. Alternatively, fresh hydrated kernels can be disrupted using stainless steel.* This method may not work as well with hydrated leaf punches or large pieces, due to there highly fibrous nature. Bead beating is the best method for processing several large samples of corn. Liquid Nitrogen, Mortar, and Pestle Method This method can be used for fresh leaf punches, kernels, root, and stem. Place a sample into the mortar or Cryogrinder well. Pour in an ample amount of liquid nitrogen to freeze the sample. Commence grinding with the pestle until ground into a fine powder. Add more liquid nitrogen throughout the process as needed. Liquid nitrogen can also be used in conjunction with the 4 ml and 15 ml vials. *Other Considerations* This method is great for sample protocols that are heat sensitive. It can be used with freshly hydrated samples Liquid nitrogen can be dangerous so take the necessary precautions when handling. Treat mortar and pestle prior to use for DNase and RNase if necessary Freeze Drying Method Freeze drying is a great way to preserve samples and a useful way to make plant tissues brittle to enhance grinding. These samples can be processed using the techniques listed above. However, with the mortar and pestle method they can be pulverized with or without treating with liquid nitrogen first, since they are already dehydrated and somewhat brittle. 96 Well Plates Seedlings and their various parts can be homogenized in 96 well formats with 5/32" stainless steel balls. Place a piece of the sample into a well. Add 100ul of buffer solution. Cap the wells with press on caps or mats and homogenize. *Other Considerations* Homogenization in a 96 well plate will generate heat.