Prac 3 answers.docx - By Joshua Saprid Determination...

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By Joshua Saprid Determination Of α-Amylase Activity In A Variety Of Seed And Seedlings Of Hordeum vulgare Questions – An Attempt 1 We began by preparing the germinating barley seeds for amylase extraction. This was achieved by drying the seeds first then their total mass recorded. The seeds were crushed to a fine paste using a mortar and pestle, then 10mL of buffer was slowly added, then crushing and mixing was continued for a further 3 minutes. The solution was then filtered into a beaker, then poured in a measuring cylinder to record the filtrate volume. After recording the filtrate was diluted five-fold by auto-pipetting 20mL of buffer into a measuring cylinder then adding 5mL of the amylase extract using the 5mL pipette to make the total volume of the solution 25mL. A control extract was created by adding 5ml of the diluted amylase extract into a separate test tube and placed into a boiling water bath for 10 minutes. After 10 minutes, the tube was removed from the bath and left to cool to room temperature. In the duration of the 10 minutes one drop of iodine was placed into each of 21 labelled ceramic wells whilst ensuring approximately equal volume for each. The reaction mixture intended to react with the iodine was created by adding 5mL of buffer and 1mL of 0.5% starch solution into separate test tube and mixed well. A plastic Pasteur pipette was utilized to add a single drop of the reaction mixture to the drop of iodine labelled T (for test) – turning the resulting solution into a blue-black hue, indicating the presence of starch. The diluted amylase extract was thoroughly remixed and 1ml of the diluted amylase extract was added to the reaction mixture and mixed well. This is the amylase reaction mixture. Starting with the well labelled

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