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.
S1
S2
S3