10X DNA LOADING BUFFER

Specifications

10X DNA Loading Buffer is widely used in nucleic acid electrophoresis. It functions as a tracking dye to monitor the migration of nucleic acids during agarose or polyacrylamide gel electrophoresis. The dyes are negatively charged and low in molecular weight, allowing them to migrate in the same direction as DNA and enabling users to visually track the movement of nucleic acid molecules through the gel. It contains three distinct tracking dyes—Bromophenol Blue, Xylene Cyanol FF, and Orange G—which provide clear visual indicators of DNA migration during electrophoresis. The presence of glycerol ensures that DNA ladders and samples form a stable layer at the bottom of the wells, preventing diffusion. Additionally, EDTA chelates divalent metal ions and inhibits metal-dependent nucleases, thereby protecting nucleic acids from enzymatic degradation.

10X DNA Loading Buffer

Manual instruction

• Use distilled water to dilute 50X TAE buffer (or 10X TBE buffer) to 1X TAE (or 1X TBE) before use.
• Weigh the appropriate amount of agarose (depending on desired gel percentage) and place it in a suitable container.
• Add the correct volume of 1X TAE (or 1X TBE) buffer and mix well.
• Boil the mixture in the microwave until the agarose is completely dissolved.
• Cool the gel to 50-55 ̊C, pour the gel into the mold and insert the comb. Be careful not to let air bubbles appear. If air bubbles appear, carefully push them to the sides with the tip of a pipette.
• Place the newly poured gel at a temperature of 4 ̊C for about 10-15 minutes (in case of urgent need) or leave it at room temperature for about 20-30 minutes, until the gel is completely solidified.
• Once cooled, place the gel tray into the electrophoresis chamber. Remove the comb and add 1X TAE (or 1X TBE) buffer to the gel tank until the buffer level is about 2 mm above the gel surface.
• Sample Loading: Load a mixture of 10X DNA Loading Buffer and sample into each well of the gel.
• Perform electrophoresis at a voltage of 85 V for 30 minutes, or until the yellow tracking dye reaches the end of the gel. The running time and voltage may be adjusted as needed based on experimental requirements.
• Observe the gel to assess DNA migration and sample separation.

Caution

• Use appropriate voltage and duration for each electrophoresis chamber.
• Ensure correct electrode placement during electrophoresis.
• Regularly clean and UV-sterilize the electrophoresis room.
• Avoid direct exposure via inhalation, skin contact, eye contact, and ingestion.
• Carefully read the instructions for use, pay attention to the expiration date of the product before use. Do not use the product after the expiration date.
• Check the packaging and product carefully before use. If the product is damaged, do not use it.
• Wear gloves, safety goggles, a face mask, and a lab coat during handling.

Read more:

• • Other Bio-reagents
  • Manual instruction