native polyacrylamide gel recipe - China Xinqi Polymer Co.,Ltd

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native polyacrylamide gel recipe - China Xinqi Polymer Co.,Ltd




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native polyacrylamide gel recipe - China Xinqi Polymer Co.,Ltd


Native polyacrylamide gel electrophoresis (PAGE) is a commonly used technique in biochemistry labs for the separation and analysis of proteins and nucleic acids. It involves the use of a polyacrylamide gel, which acts as a sieving matrix to separate molecules based on their size and charge. While commercially available precast gels are convenient, they can be expensive and may not always meet the specific needs of a particular experiment. cationic polymer coagulant While commercially available precast gels are convenient, they can be expensive and may not always meet the specific needs of a particular experiment. In such cases, a native polyacrylamide gel recipe can be a useful tool for researchers to create their own customized gels. Here is a step-by-step guide to creating a native polyacrylamide gel using a simple recipe. Ingredients: - Acrylamide powder - Bis-acrylamide powder - Tris buffer - Ammonium persulfate (APS) - TEMED - Distilled water - Gel casting tray - Comb Step 1: Prepare the gel casting tray Start by cleaning the gel casting tray with soap and water and then rinse with distilled water. Dry the tray with a lint-free cloth to avoid any contamination. Place the tray on a level surface and insert the appropriate comb to create the desired number and size of wells. Step 2: Prepare the gel solution In a clean beaker, mix the following ingredients: - 30% acrylamide solution - 10% bis-acrylamide solution - 1M Tris buffer (pH 8.8) - Distilled water The volume of each ingredient will depend on the size of the gel you want to make. A general rule of thumb is to use a 29:1 ratio of acrylamide to bis-acrylamide for a 10% gel and a 37.5:1 ratio for a 12% gel. The final volume of the gel solution should be 10-15% more than the volume of the gel casting tray. Step 3: Add the initiator and catalyst To polymerize the gel, you will need to add two important components, ammonium persulfate (APS) and TEMED. These chemicals work together to initiate the polymerization process. Add 10% of 10% APS solution and 0.1% of TEMED to the gel solution and mix well. Step 4: Pour the gel Carefully pour the gel solution into the gel casting tray, making sure to avoid any air bubbles. The gel should be poured to the top of the tray, leaving some room for the stacking gel if needed. Insert the comb and let the gel polymerize for 30-45 minutes. Step 5: Remove the comb and add the stacking gel (optional) If you want to separate proteins or nucleic acids of similar size, you can add a stacking gel on top of the separating gel. To prepare the stacking gel, follow the same steps as for the separating gel, but with a lower concentration of acrylamide (usually 4-5%). Once the separating gel has polymerized, remove the comb and pour the stacking gel on top. Let it polymerize for an additional 30 minutes. Step 6: Prepare the gel for electrophoresis Once the gel has polymerized, carefully remove it from the casting tray and place it in the electrophoresis chamber. Fill the chamber with 1X Tris-glycine running buffer, making sure the gel is completely submerged. Step 7: Load the samples and run the gel Add your protein or nucleic acid samples to the wells using a micropipette and a loading dye. The loading dye helps to visualize the sample as it moves through the gel. Connect the electrodes to the power supply and run the gel at a constant voltage. Step 8: Stain and visualize the gel Once the samples have migrated through the gel, remove it from the chamber and carefully remove the comb. Stain the gel with a protein or nucleic acid stain, depending on your experiment, and visualize the separated bands under UV light or by using a gel imaging system. Congratulations, you have successfully created a native polyacrylamide gel using a simple recipe! This gel can be used for a variety of experiments, such as protein purification, identification of protein-protein interactions, and DNA sequencing. By making your own gel, you have the flexibility to customize it according to your specific experimental needs, saving time and money in the long run. In conclusion, using a native polyacrylamide gel recipe is a cost-effective and efficient way to create customized gels for your biochemistry experiments. With just a few simple ingredients and steps, you can create a gel that meets your specific requirements and produces reliable results. So next time you need a polyacrylamide gel, consider making your own using this easy-to-follow recipe.