I have a question:( refrence plant cell culture Michel Davey ...] does any body know why in this example of making 2oo ml of EMS it is said to reach the volume to 195.5 ? i mean (1ml EMS +4 ml DMSO + 195 ml water = 2oo ml final volume while it has been said to add 195.5 ml water to the mixed please help me Preparation of EMS solution: 1 Use only freshly prepared EMS solution. This consists of EMS, the active ingredient, DMSO as the carrier agent and distilled water. Prepare the EMS solution in two phases: First, mix the required volumes of water and 2% (v/v) DMSO and autoclave at 120 ◦C for 15 min at 103.5 kPa (15 psi). Leave the mixture to cool to room temperature. This step may carried out in advance, and the sterile mixture used for the preparation of the EMS solution up to 24 h later. The second phase, which must be carried out in a laminar flow cabinet, involves the addition of EMS to the water–DMSO mixture. When ready to incubate the target materials in the mutagen, use a sterile syringe and a 0.2 μm filter to add the required volume of EMS solution to the sterile water–DMSO mixture. Shake the resulting solution vigorously to give an homogeneous emulsion. Example: To prepare 200 ml of 0.5% (v/v) EMS with 2% DMSO, mix 4 ml of DMSO and 1ml of EMS solution (Sigma, d = 1.17 g/ml) in 195.5 ml distilled water. As a guide for the volume required, prepare 1 ml of solution for every seed to be treated. Pretreatment handling of seeds and determination of optimal treatment conditions
Posted on: Fri, 28 Jun 2013 09:17:44 +0000