Southern California Biotechnology Center (SCBC) at Miramar College: PCR ALU
Standard Operating Procedure (SOP): December, 2015
Title: Preparing 50X Gel Green Loading Dye and Marker for PCR ALU Lab
Materials/Supplies:
10,000X Gel Green Nucleic Acid stain (EmbiTec Cat#EC-1995)
Orange G (use the “Preparing Orange G” Protocol)
Amber microfuge tubes
Micropipette
Pipette tips (sterile)
Prepare 1.0 ml of 50X Gel Green in Orange G from a 10,000X stock of Gel Green
1. Obtain the 10,000X gel green stock solution (stored in an amber conical tube on
the Amgen Shelf).
2. Using a p-20 micropipette, aseptically remove 5µl of gel green from the original
stock tube and dispense into a new amber microfuge tube labeled “50X gel
green.”
3. Using a p-1000 micropipette, add 995 µl of Orange G loading dye to this tube.
4. Label amber microfuge tubes with “LD.”
5. Aliquot 140 µl of gel green loading dye per tube for Amgen lab 4
6. Aliquot 60 µl of gel green loading dye per tube for Amgen lab 4a
7. Store at room temperature.
Prepare the DNA Marker as below:
1. Label an amber microfuge tube “M”
2. Prepare the 1 kb plus DNA marker by performing a 1:25 dilution of the marker
using the 50X Gel Green loading dye prepared above.
3. Example: 10 ul of DNA marker + 240 ul of 50X gel green loading dye.
4. Aliquot 130 µl of DNA Marker per tube for Amgen labs 4 and 4a
5. Store at room temperature.
*** Note: Gel green and Marker should be stored at room temperature, protected from
light! At lower temperatures, precipitation of the dye may occur. Also, Prepare the Gel
green solution and the marker at the time the kit is going out. For best results, it should
be used within 2 weeks of being prepared.