SYBR Safe DNA Stain is a quick and easy method to visualize DNA in agarose gels. It is a safer and more sensitive DNA stain alternative to ethidium bromide, and can be used with blue light or UV exposure. This stain can be found in many of the Edvo-Kits, most commonly in the: Ready-to-Load DNA Electrophoresis kits, Polymerase Chain Reaction (PCR) kits, and Advanced DNA Application kits. Given the major role of SYBR Safe in DNA visualization, here is everything that you need to know about this stain!
Dilute! Dilute! Dilute!
Why do our protocols call for SYBR dilutions? Well, that small amber tube that you receive with SYBR in it is actually very concentrated. So, by diluting it with distilled water or electrophoresis buffer we can make it less concentrated while also making the addition of SYBR into agarose gels easier.
A Quick Overview of How to Dilute SYBR
While the dilution scheme and volumes/types of reagents can vary slightly from kit to kit, the main steps remain the same. The following is an example of the steps taken to dilute SYBR in Edvo-Kit 330: PCR Amplification of DNA.
- Prepare 1x Electrophoresis Buffer by combining 10 uL of 50x Concentrated Buffer with 490 uL of distilled water.
- If the aliquot in the amber tube is not all present at the bottom of the tube, centrifuge the tube for a few seconds before adding the buffer.
- Add 390 uL of 1x Electrophoresis Buffer from step 1 to the tube of SYBR safe and mix by tapping the tube several times. This is your diluted SYBR, which is now ready to be used during your agarose gel preparation.
- After microwaving your agarose solution, cool the agarose to 60°C by swirling often to dissipate the heat.
- Following the table in your literature, add diluted SYBR to the molten agarose. For example: if you are casting a 7×7 cm gel, add 30 uL of diluted SYBR to 30 mL of molten agarose.
- Let the molten agarose with diluted SYBR cool in the casting tray.
- The gel is now ready to be used.
Remember, look at the literature for each kit for the instructions on how to dilute SYBR Safe DNA Stain, as it often varies slightly from kit to kit.
Frequently Asked Questions
Q: The amber tube of SYBR Safe DNA Stain I received looks empty, what should I do?
A: We send a small aliquot of SYBR because it is very concentrated. Centrifuging the tube for a few seconds will help collect it at the bottom, making it easier to dilute.
Q: Where can I find the instructions for diluting SYBR Safe DNA Stain in the literature?
A: It can be found in the Instructor’s Guide section of the literature in a bright green box. It is usually on the same page where everything about preparing agarose gels and electrophoresis buffer has been outlined.
Q: How much diluted SYBR Safe DNA Stain do I add per gel?
A: In the student protocol, there will be a section about preparing agarose gel with diluted SYBR Safe. On this page, there is a table that outlines how much diluted stain to add based on the size of the gel casting tray and the volume of agarose being used.
Q: I am using the Melt and Pour UltraSpec-Agarose, can I still use SYBR Safe stain to visualize DNA?
A: Yes, the instructions for using SYBR will be the same. Heat the Melt and Pour UltraSpec-Agarose in the microwave, let it cool briefly, and add the diluted SYBR Safe using the table in the student protocol as a guide.
Q: What is the difference between using FlashBlue and SYBR Safe DNA Stain?
A: They are two different methods used to visualize DNA in agarose gels. FlashBlue requires a white light system to visualize DNA, while SYBR requires blue light or UV exposure. If you are using both, remember to visualize the SYBR Safe bands before staining with the FlashBlue.
Please reach out to an Edvotek® Technical Support Specialist at 1-800-EDVOTEK or email us at firstname.lastname@example.org if you have any questions.