Gel electrophoresis operation precautions

1. Use high-quality agarose: different brands of agarose have a big difference in quality. Inferior agarose can cause DNA bands to be blurred and even bands missing. Therefore, please try to choose a brand with stable quality.

2. Choose the appropriate gel concentration:

Linear DNA resolution range of agarose gel

3. Gel preparation: The same 1x buffer should be used for gel and electrophoresis; the volume of the container used is at least 3 times that of the gel solution to avoid overflow when the solution boils; the bubbles are removed as much as possible during the gel making process; Select the appropriate size comb for the volume.

4. Select the appropriate electrophoresis buffer: GenStar® SD Buffer (Cat. No. E101-50) and TBE buffer for separation of relatively small DNA fragments, TAE buffer (Cat. No. E102-50) for separation and comparison Large DNA fragments; buffers should be updated in a timely manner; freshly prepared 1x running buffer should be used for gel recovery to prevent nuclease and DNA band contamination.

5. Preparation of nucleic acid samples: Extracted DNA samples should be removed from impurities such as proteins and excess salts; PCR samples should be detected by electrophoresis within 24 hours, otherwise the bands are easily blurred.

6. Loading Buffer: The same loading buffer should be used for all lanes to prevent band mobility deviation due to differences in ionic strength.

7. Appropriate loading: too much loading will cause the loading hole to be overloaded, and the towing phenomenon will occur; if the loading volume is too small, the brightness of the strip may be uneven.

8. Voltage and electrophoresis time: Select the appropriate voltage and electrophoresis time according to the type of electrophoresis tank, gel concentration, type of electrophoresis buffer, and size of the target strip. When using GenStar® SD running buffer, the voltage is set to 20~35 v/cm gel length; when using TAE or TBE buffer, the maximum voltage is not more than 20 v/cm.