I have isolated good quality DNA from plant using CTAB method. when i diluted these DNA (20 ng/ul) the DNA get degraded. I have check the diluted DNA on 1% gel but i dont seen the DNA.
DNA degradation during dilutions can be a concern, especially if the dilution process involves extended exposure to environmental factors or if the DNA is particularly sensitive.
1. Ensure that your work area is clean and free from contaminants. Use appropriate laboratory practices, including wearing gloves and using sterile equipment.
2. Use molecular biology-grade water or (MilliQwater) and reagents that are certified as RNase-free to minimize the risk of introducing nucleases that could degrade DNA.
3. Store DNA samples at an appropriate temperature and avoid freeze-thaw cycles, as repeated freezing and thawing can lead to DNA degradation. Use a dedicated freezer for long-term storage.
4. Work with the smallest volume possible for your dilutions to minimize the time DNA spends in the dilution buffer. If necessary, optimize the dilution buffer composition to provide optimal conditions for DNA stability.
5. Choose low-binding tubes and pipette tips to minimize the loss of DNA due to adherence to plastic surfaces. This is particularly important when working with low DNA concentrations.
6. If possible, perform dilutions on ice or in a refrigerated environment. Lower temperatures can slow down enzymatic activity and reduce the risk of DNA degradation.
7. Consider using dilution buffers that contain DNA stabilizers, such as EDTA, to protect DNA from nucleases and other degrading factors and Minimize the time DNA spends in the dilution buffer. Plan your dilution process carefully and work efficiently.
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