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12 Dec 2019

I need help doing problem number 4 listed below. If someone could show the work required for this problem it would be a great help!

Below is a list of solutions you will need for the mini-prep. Make 5mL of each solution as a team and store each in a 15mL Falcon tube. Store solution 1 and 70% ethanol at 4 degrees Celsius. Solution 3 and Elution Buffer can be stored at room temperature. Show your calculations in the space provided below.

1. Solution 1 (lysis buffer) is (25mM Tris-HCl, pH 8.0, 50mM glucose, 10mM EDTA)

✓ Recall that you have Tris and EDTA stock solutions

✓ You will be provided with a 3M glucose stock solution

2. Solution 2 (Denaturing solution) is (0.2N NaOH, 1.0% SDS)

✓ Do NOT make today (it needs to be made fresh on the day of the mini-prep)

✓ You will be provided stocks of 1N NaOH and 10% SDS

3. Solution 3 (Renaturing Solution) is 5M potassium acetate.

✓ Add 3mL of 5M potassium acetate to 0.575mL of glacial acetic acid and 1.425mL of water

4. 70% ethanol (v/v)

✓ You will be provided with 100% (200 proof) ethanol

5. Elution Buffer (EB) is (10mM Tris-HCl, pH 8.0)

✓ Recall that you have Tris stock solution

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Irving Heathcote
Irving HeathcoteLv2
13 Dec 2019

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