Team:Northwestern/Protocols/Solutions

Materials:

• 10 mM KOAc pH 7.0 (10mL of 1M stock/L)
• 80 mM CaCl2 x 2 H2O (11.8 g/L)
• 20 mM MnCl2 x 4 H2O (4.0 g/L)
• 10 mM MgCl2 x 6 H2O (2.0 g/L)
• 10% Glycerol (100 mL/L)

Notes:

• Adjusting pH up may precipitate MnO2 from the Mn containing solutions
• Slight dark precipitate will not impede function of solution

Materials:

• 10 g Tryptone
• 5 g Yeast Extract
• 10 g NaCl
• 10 g of Agar (for 1% Agar Plates (w/v))
• 1 L ddH2O
• Antibiotic Stock (if Required)

Notes:

• Do not store, the gel will condense and become unusable
• Add the Antibiotic stock after pouring some non-antibiotic LB Plates
• 1 L Should pour ~40 Plates

Materials:

• 0.5% (w/v) Yeast Extract (5 g/L)
• 2% (w/v) Tryptone (20 g/L)
• 10 mM NaCl (.584 g/L)
• 2.5 mM KCl (.186 g/L)
• 20 mM MgSO4 (2.4 g/L)

Notes:

• Autoclave and Store at 4C

To make 1 liter of 5X TBE buffer:

1. Mix 54 g Tris (Trizma base) and 27.5 g Boric Acid in 900 mL DI water in a 2 L Erlenmeyer flask.
2. Add 20 mL 0.5M EDTA solution.
3. Pour into a 1 L graduated cylinder.
4. Adjust volume of solution to 1 L with DI water.
5. Store at room temperature.

Making 100 mL of 0.5M EDTA solution:

1. Add 18.61 g EDTA to 100 mL DI water.
2. The EDTA will not dissolve unless the solution is at a pH of 8. Bring the solution to the biology labs to use the pH meter. Follow the instructions by the wall next to the pH meter for calibration, etc.
3. Place the solution on a stirrer and add a magnetic stir bar.
4. Adjust the pH of the solution with “Sat. NaOH” (saturated NaOH). Use a pipette from the drawers. The pH will go up, peak, then go down. Add more NaOH as this happens. If the “A” has stopped blinking on the meter, it has stopped acquiring. Press “Read” to start acquiring again.
5. Repeat until solution is clear and pH is stable at pH 8.