Southern California Biotechnology Center (SCBC) at Miramar College: Amgen Biotech Experience(ABE) Program

Standard Operating Procedure (SOP): December, 2015

Title: Preparing 20X Sodium Borate (SB) Buffer -1 Liter for Amgen Labs 1 and 4/4a.

Materials/Supplies:

Boric Acid

Sodium Hydroxide(NaOH)

DI water (dH20)

Balance

Spatula

Stir plate

Stir bar (rinse with dH2O)

pH Meter

Graduated cylinder (1 liter) or larger

Beaker (1 liter) or larger

Storage bottle (1 literor larger)

Tape

Sharpie

Procedure for preparing 20X SB buffer:

Caution: Do not add NaOH and Boric Acid to the water at the same time. They will clump together and take very long to go into solution!

  1. Obtain materials and supplies listed above.
  2. Place stir bar in beaker.
  3. Weigh out 8.0 grams of Sodium Hydroxide (NaOH) place in beaker.
  4. Add 800 ml dH20 to beaker place on stir plate and stir until NaOH dissolves.
  5. Weigh 48 grams of Boric Acid.
  6. Add Boric Acid to NaOH solution stir to dissolve.
  7. Check pH of solution.
  8. Adjust pH to 8.0 using concentrated Boric Acid or concentrated NaOH solution.
  9. Pour solution back into the empty 1 liter graduated cylinder and add dH2O to a volume of 1 liter.
  10. Pour 20X SB solution into a 1 liter bottle cap tightly.
  11. Use a piece of tape and a sharpie to write the contents of the bottle, the pH, date prepared and who prepared by and needs QC testing.
  12. Store at room temperature.

Procedure for preparing 1 liter of 1X SB buffer:

  1. Add 50mL of 20X SB buffer to 950mL of dH2O.
  2. Place in 1 liter bottle.
  3. Use a piece of tape to label bottle with contents, pH, date prepared and who prepared by.

Procedure for quality testing of 1X SB buffer:

  1. Use the protocol for Amgen lab 1 in this binder to prepare a 0.8% agar or agarose gel and perform Amgen lab 1.
  2. Results should be as shown on the photo below.
  3. Once buffer has passed QC please label the 20x bottle with date of QC testing and whether it passed or not.