AB + AF 1xMBSH Recipe

AB + AF and 1x MBSH

-Used for oocyte storage solution (Fridge C)-

Recipe:

• 1mL of AB + AF

• 100ML of 1x MBSH

= Makes 100mL of AB + AF and 1x MBSH

(Read Before)

Notes:

AB + AF vials are 100x, and so need to be diluted 100-fold.

The vials are ~1mL (milliliters) in volume. The pipetters are set to pull in μL (microliters).

μL =/= mL

1000μL = 1mL

One 1mL of AB + AF will dilute into 100mL of 1xMBSH making 100mL of the AB + AF 1x MBSH solution. This is the solution that the oocytes are kept in.

• You must be pipette trained before making this solution!

Directions:

1. Grab a capsule of AB + AF from Freezer B. It should be in a box labeled "AB + AF".

2. Grab the 1xMBSH solution from Fridge A, NOT the 10x MBSH.

3. Grab a pipette, one that's 100/200μL - 1000/2000 μL, and the matching pipette tips.

4. Grab colored tape and a sharpie.

5. Put on gloves and then sterilize hands with Eliminase.

6. Grab a clean, autoclaved, bottle that is a little larger than the volume you're needing, but not too large.

• For example, use a 200mL bottle if you're needing 100mL, 150mL, or 200mL. Do not use a 750mL bottle if you only need 100mL of solution.

7. Take all supplies over to the Equipment Room 515 to work in the hood. Turn on the ventilation.

8. Pour the 1x MBSH into the new bottle, to the desired volume.

• Note: If you're using a full capsule of AB + AF, you need to use at least 100mL of 1x MBSH to get the correct dilution.

9. Close the 1xMBSH bottle and set aside.

10. Ready the pipette by setting the desired volume. [Remember that this reading is in microliters. There are 1000 microliters in one milliliter].

11. Pipette the desired amount into the solution bottle.

12. Close the lid on the new solution bottle and mix.

13. With the tape and a sharpie, label the bottle with your initials, the current date, and the name of the solution / concentration. Write legibly and large enough so that others can read it.

14. Return all of your materials to their correct locations, and turn off the ventilation in the hood.

15. Leave Room 515 after turning off the lights and hood.

16. Place the new AB + AF 1x MBSH solution in Fridge A.