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5 September, 14:14

During the Purification of Lactate Dehydrogenase (LDH) experiment, you will need 50ml of buffer A150. Buffer A150 is 30mM Tris (pH 8.8) and 150mM NaCl. Given 1L of 1.5M Tris and 500ml of 5M NaCl, how much of each stock do you need to make 500ml of the A150 buffer?

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  1. 5 September, 17:50
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    You will need 10 mL of stock Tris, 15mL of stock NaCl and enough water to bring the total volume to 500 mL.

    Explanation:

    The desired buffer contains:

    Total volume of 500 mL Tris 30mM NaCl 150mM

    The stock solutions are:

    1.5M = 1500mM Tris 5M = 5000mM NaCl

    We can use the formula C1V1=C2V2 to calculate the volume of each stock required to prepare the buffer.

    Tris C1 = 1500 mM V1=? C2 = 30mM V2 = 500 mL

    1500 mM x V1 = 30mM x 500 mL

    V1 = (30mM x 500 mL) / 1500 mM

    V1 = 10 mL

    NaCl C1 = 5000 mM V1=? C2 = 150 mM v2 = 500 mL

    5000 mM x V1 = 150mM x 500 mL

    V1 = (150 mM x 500 mL) / 5000 mM

    V1 = 15 mL

    You will need 10 mL of stock Tris, 15mL of stock NaCl and enough water to bring the total volume to 500 mL.
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