DNA Agarose Nucleic Acid Electrophoresis

Summary

Nucleic acid electrophoresis is an important tool for conducting nucleic acid research and can be used for techniques such as (1) nucleic acid probing; (2) nucleic acid amplification; and (3) sequence analysis.

Operation method

DNA agarose nucleic acid electrophoresis

Principle

The tendential movement of a charged substance in an electric field is called electrophoresis. Nucleic acid electrophoresis is an important tool for nucleic acid research and is an indispensable component of techniques such as nucleic acid probes, nucleic acid amplification and sequence analysis. Nucleic acid electrophoresis is usually carried out in agarose or polyacrylamide gels. Agarose and polyacrylamide at different concentrations can form gels with different molecular sieve mesh sizes, which can be used to separate nucleic acid fragments of different molecular weights.

Materials and Instruments

DNA Samples
Agarose dry powder Electrophoresis buffer Sample loading buffer
Triangle flasks Microwave oven Electrophoresis apparatus Electrophoresis tanks

Move

1. Rinse the gel-making mold and comb with distilled water, place them on the gel-making plate, close the edges of the mold, and set up the comb;

2. According to the size of the DNA fragments to be separated, prepare agarose gel with gel buffer of appropriate concentration: accurately weigh the dry agarose powder, add it to the triangular flask used for gel preparation, and quantitatively add electrophoresis buffer (generally 20~30 ml);

3. Put it into the microwave oven and melt it. Cool down for a while, add a drop of fluorescent dye, gently rotate to mix the gel solution well, pour it into the electrophoresis tank, and wait for it to solidify. 4;

4. After the gel has completely solidified for 30~45 minutes at room temperature, pull out the comb carefully and place the gel in the electrophoresis tank;

5. Pour electrophoresis buffer into the electrophoresis tank, the amount of buffer should be 1mm above the surface of the gel, if there are air bubbles in the sample wells, they should be removed;

6. Add 10×volume of loading buffer into the DNA sample, mix well, and then slowly add the sample mixture into the submerged gel sample wells with a gun;

7. Turn on the power, red color is positive pole, black color is negative pole, remember that the DNA sample swims from negative pole to positive pole (the end close to the filling hole is negative). Generally 60-100V, electrophoresis for 20-40min is enough. 8;

8. Judge whether to terminate the electrophoresis according to the position of the indicator;

9. When electrophoresis is finished, turn off the power, observe the electrophoretic bands and their positions on the gel imager, and compare the size of the amplified products with the standard marker of nucleic acid molecular weight.

Caveat

When making glue, you need to melt the glue evenly to avoid affecting the results of the subsequent running glue.

Common Problems

In addition to agarose nucleic acid electrophoresis, polyacrylamide gel electrophoresis can be performed, the latter being suitable for the separation of oligonucleic acids.

For more product details, please visit Aladdin Scientific website.

https://www.aladdinsci.com/