Some Practical Considerations
1. It's important to prepare the electrophoresis buffer and gel fresh each time. The cutting table should be thoroughly cleaned, and the blade must be sterilized to avoid any contamination. This step is crucial in ensuring that the cut gel piece remains free from foreign DNA, which could interfere with your results.
2. When excising the gel, make sure to collect all of the gel containing the target DNA fragment. To minimize the volume, use a thin layer of gel, as long as it’s sufficient for recovery. A thin and wide comb can also help in running the gel more efficiently, especially when working with smaller samples.
3. For personal protection, wearing regular plexiglass shields is usually enough. However, it’s recommended to wear a protective mask to safeguard your eyes, especially if you are using resin glasses or working in an environment where UV exposure might be a concern.
If you're particularly sensitive to UV light or have specific requirements—such as needing an EB-free procedure—you can use a marker and a ruler to take photos under UV light. This will help you accurately locate the target band on the gel.
Follow the standard procedure: mark the gel, cut out the desired region using a marker, stain with EB, and take a photo with a ruler placed under the UV lamp. Since the position of the marker relative to the DNA band is known, you can measure the exact location of the target band on the unstained gel.
If you find it difficult to estimate the position, you can lightly mark a small dot directly on the marker. This makes it much easier to identify the correct area when cutting the gel. Always double-check your measurements before proceeding to ensure accuracy and prevent unnecessary loss of valuable DNA samples.
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