Preparing an Agarose Gel For Electrophoresis

Preparation of Agarose Gel for Electrophoresis

Note: Always wear disposable gloves!

Step-by-Step Instructions

Weighing Agarose Powder: use weighing paper and an electronic balance to measure 0.3g of agarose powder (based on a 30ml system).

Preparing TBST Buffer: prepare 30ml of 1x TBST buffer in a 100ml Erlenmeyer flask.

Dissolving Agarose Powder: pour the agarose powder into the TBST buffer and shake well. Put them into a microwave and heat (typically for 50 seconds, covered with aluminum foil) until completely dissolved.

Cooling and Adding Nuclease:use gloves to remove the mixture from the microwave and let it cool slightly in cold water until it is warm (approximately 60°C). Add 2µl of nuclease (Eb substitute) while shaking to mix thoroughly.

Preparing the Gel Mold:

• Clean and dry the gel tray and gel casting device of the electrophoresis tank.
•  Place the gel tray into the inner tank and insert the comb at a fixed position.
• Mix the agarose gel solution, cooled to about 65°C, and carefully pour it onto the gel tray in the gel casting device, spreading it slowly until it forms an even gel layer on the glass plate.
•  Allow it to sit at room temperature until the gel solidifies completely.
• Gently remove the comb vertically and remove the tape.
• Place the gel tray into the electrophoresis tank.

Important: Ensure there are no air bubbles at the comb teeth area. The surface of the liquid should be smooth without ripples.

Running the Gel

Loading the Gel

After the gel solidifies, place it into the electrophoresis tank and pour the electrophoresis buffer until the gel is submerged.

Preparing Samples

•  Take the marker and loading buffer from the refrigerator.
•  Add 6µl of loading buffer to the samples and mix well.
• Using a micropipette, slowly load the samples into the large wells of the gel (be careful not to puncture the gel and do not dispense the entire volume to avoid air bubbles).
•  Load the marker into any of the small wells (remember its position).

Starting Electrophoresis

• Cover the electrophoresis tank and start electrophoresis immediately after loading the gel.
• Set the voltage to 60-100V. The samples will migrate from the negative electrode (black) to the positive electrode (red).
• Higher voltage shortens the effective separation range of the agarose gel.
• Stop electrophoresis when the bromophenol blue dye reaches about 1cm from the bottom edge of the gel plate.

Observing Results

After separating the bands, stop electrophoresis, take out the gel, and directly detect and photograph it.

Use a gel imaging system to photograph and observe the bands (check if there are any bands for the marker or samples).

After obtaining your gel band map, find the marker. Based on the marker, you can determine the target bands!

Beijing Liuyi Biotechnology Co. Ltd (Liuyi Biotechnology) have been manufactured electrophoresis products for more than 50 years. We present our horizontal electrophoresis systems here for agarose gel electrophoresis. Choose horizontal electrophoresis systems from Beijing liuyi for your gel electrophoresis experiment to help your research in biotechnology industry.

We offer different models of horizontal electrophoresis tanks for casting and running small agarose gels to big agarose gels

Selection Guide for Horizontal Immersed Gel Electrophoresis Unit

Beijing Liuyi Biotechnology Co. Ltd (Liuyi Biotechnology) has specialized in manufacturing electrophoresis instruments for more than 50 years with our own professional technical team and R&D center. We have reliable and complete production line from design to inspection, and warehouse, as well as marketing support. Our main products are Electrophoresis Cell (tank/chamber), Electrophoresis Power Supply, Blue LED Transilluminator, UV Transilluminator, Gel Image & Analysis System etc. We also supply the lab instruments like PCR instrument, vortex mixer and centrifuge for laboratory.

If you have any purchasing plan for our products, please don’t hesitate to contact us. You can send us message at email [email protected] or [email protected], or please call us at +86 15810650221 or add Whatsapp +86 15810650221, or Wechat: 15810650221.

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