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Preperation of Trypsin-EDTA Solution

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  • Preperation of Trypsin-EDTA Solution

    Hi,

    I'm trying to prepare trypsin-EDTA solution and would like some help with the math.

    I have 2.5% trypsin in 500ml and 0.5M EDTA.

    How do I go about preparing the final dissociation composition of 0.25%Trypsin-0.03% EDTA in final volume of 50ml?

  • #2
    Originally posted by MsT View Post
    Hi,

    I'm trying to prepare trypsin-EDTA solution and would like some help with the math.

    I have 2.5% trypsin in 500ml and 0.5M EDTA.

    How do I go about preparing the final dissociation composition of 0.25%Trypsin-0.03% EDTA in final volume of 50ml?
    Per Wikipedia, molar mass of EDTA is 292.24 g/mol, so 0.5M is 146.12 g/L. I assume 0.03% is 300 mg/L

    For each component, C1*V1 = C2*V2, use an amount of stock V1 = V2*C2/C1, where V2 is 50 mL

    Trypsin: V1 = 50 mL *0.25%/2.5% = 5 mL
    EDTA V1 = 50 mL * 300 mg/L / 146.12 g/L = 62 L
    Dilute to final volume of 50 mL

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    • #3
      Thank you for your reply. That was really useful. I have another question. I have 1MG of IFN-y powder (mw- 17145.6 g/mol) and I want to reconstitute in dH2O to have a stock concentration of 0.5 mg/mL. What's the volume of dH2O required for the reconstitution.

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      • #4
        Originally posted by MsT View Post
        Thank you for your reply. That was really useful. I have another question. I have 1MG of IFN-y powder (mw- 17145.6 g/mol) and I want to reconstitute in dH2O to have a stock concentration of 0.5 mg/mL. What's the volume of dH2O required for the reconstitution.
        I assume you mean 1 mg (case is important in metric). If you want 0.5 mg/mL, you only have enough to make up 2 mL of it. Mix with a little less than 2 mL of water, then bring to final volume of exactly 2 mL

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        • #5
          Thank you!

          To further dilute this to a 200ug/50ul:

          v1 x c1=v2 x c2
          V1 x 500ug/mL = 50ul x 200ug
          V1= 20ul

          So 20ul of stock in 30ul of water.

          Is this calculation right?

          Comment


          • #6
            Originally posted by MsT View Post
            Thank you!

            To further dilute this to a 200ug/50ul:

            v1 x c1=v2 x c2
            V1 x 500ug/mL = 50ul x 200ug
            V1= 20ul

            So 20ul of stock in 30ul of water.

            Is this calculation right?
            Hi MsT, there are 1000L in 1mL.

            You have a solution that is 500g in 1000L, you want 200g in 50L. That is not a dilution, you have to make it more concentrated.

            Have you considered whether there are any safety issues around you trying to prepare these solutions? - For your own sake, is there a supervisor or mentor helping?

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