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

Standard Operating Procedure (SOP): December, 2015

Title: Preparing Dyes for Amgen Lab 1

Materials/Supplies:

Scale

Spatula

Weigh paper or trough

Graduated cylinder

15 ml or 50 ml conical tubes

Glycerol

Orange G

Bromophenol Blue

Xylene Cyanol

1. Prepare glycerol (you will need a total volume of 30 ml to prepare solutions 1, 2 and 3) by diluting 100% glycerol solution to 50% glycerol as below.

Note: Glycerol is very viscous so be patient when working with it.

32 ml of 50% v/v glycerol = 16 ml of 100% glycerol + 16 ml of DI H20.

Prepare solutions 1, 2 and 3 as below:

Solution 1

15.0 mg of Bromophenol Blue

10 ml of 50% glycerol

10 ml of deionized water

Vortex to dissolve

Solution 2

15.0 mg of Bromophenol Blue

10 ml of 50% glycerol

10 ml of deionized water

15.0 mg of Xylene Cyanol

35.0 mg of Orange G

Vortex to dissolve

Solution 3

10 ml of 50% glycerol

10 ml of deionized water

15.0 mg of Xylene Cyanol

Vortex to dissolve

To prepare class sets of dyes, aliquots the dyes into colored and numbered eppendorph tubes as below.

Solution 1: Aliquot 0.5ml of solution into YELLOW eppendorph tubes. Close tubes and use a sharpie to write the number “S1” on the caps of each tube. Place tubes in a styrofoam box.

Solutions 2: Aliquot 0.5ml of solution into GREEN eppendorph tubes. Close tubes and use a sharpie to write the number “S2” on the caps of each tube. Place tubes in a styrofoam box.

Solutions 2: Aliquot 0.5ml of solution into BLUE eppendorph tubes. Close tubes and use a sharpie to write the number “S3” on the caps of each tube. Place tubes in a styrofoam box.

Label the styrofoam box with contents, date prepared and who prepared by. Store in fridge or cold room.

Procedure for quality testing of Dyes:

1.  Use the protocol for Amgen lab 1: Preparing 0.8% agar or agarose gels in this binder to prepare a 0.8% agarose gel and perform Amgen lab 1.

2.  Results should be as shown on the attached photo.

3.  Once dyes have passed QC label the styrofoam box with date of QC testing and whether it passed or not.