Corn seed purity identification reagent preparation method
Method for Preparing Reagents for Corn Seed Purity Identification
First, prepare the electrode buffer. Weigh 6.00 g of glycine and place it into a 2000 ml beaker. Add approximately 1800 ml of deionized water and dissolve the glycine. Adjust the pH to 3.3 using about 2.0 ml of lactic acid. Then add deionized water to reach a total volume of 2000 ml and mix thoroughly.
Second, prepare the sample extract. Weigh 5.80 g of sodium chloride, 200 g of sucrose, and 0.15 g of methyl green, and place them in a 1000 ml beaker. Add approximately 800 ml of deionized water and dissolve the mixture. Heat the solution to a gentle boil, then allow it to cool. Finally, add deionized water to bring the total volume to 1000 ml. Store the solution in a refrigerator at 4°C.
Third, prepare the separation gel buffer. In a 1000 ml beaker, measure 1.43 ml of sodium lactate and add approximately 980 ml of deionized water. Adjust the pH to 3.0 using lactic acid, and then dilute to a final volume of 1000 ml. Transfer the solution to a brown bottle and store it in a refrigerator at 4°C.
Fourth, prepare the separation gel solution. Weigh 112.5 g of acrylamide, 3.75 g of N,N'-methylene bisacrylamide, 0.25 g of ascorbic acid, and 8.0 mg of ferrous sulfate. Dissolve these components in the previously prepared separation gel buffer, and adjust the volume to 1000 ml. Filter the solution and store it in a brown bottle in a refrigerator at 4°C (use within 2 weeks).
Fifth, prepare the concentrated gel buffer. In a 200 ml beaker, measure 0.30 ml of sodium lactate and add approximately 90 ml of deionized water. Adjust the pH to 5.2 with lactic acid, and bring the volume to 100 ml. Store the solution in a brown bottle and keep it in a refrigerator at 4°C.
Sixth, prepare the concentrated gel solution. Weigh 6.00 g of acrylamide, 1.00 g of N,N'-methylene bisacrylamide, 0.03 g of ascorbic acid, and 0.8 mg of ferrous sulfate. Dissolve these ingredients in the concentrated gel buffer and adjust the volume to 100 ml. Store the solution in a brown bottle and keep it in a refrigerator at 4°C (use within 1 week).
Seventh, prepare a 3% hydrogen peroxide solution. Take 1 ml of 30% hydrogen peroxide and mix it with 9 ml of deionized water. Store the solution in a brown bottle and refrigerate at 4°C.
Eighth, prepare the staining solution. Weigh 2.00 g of Coomassie Brilliant Blue R250 and grind it with 100 ml of anhydrous ethanol in a mortar and pestle. Filter the solution and transfer it to a brown bottle. Take 10 ml of this solution and add it to 200 ml of 10% (w/v) trichloroacetic acid solution. Mix well before use.
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