according to protocol that i am following in which i am using tris edta for washing, my dna samples are not transparent instead these are some creamy and also during running on gel electrophoresis bands are not clear.
Difference between Tris-EDTA TE Buffer and Washing Buffer:
Washing Buffer
Washing buffers are usually used to remove unwanted contaminants from the DNA preparation. These buffers often contain salts (like sodium chloride), alcohol (ethanol or isopropanol), or detergents. They help remove proteins, lipids, and other cellular debris from the sample after the DNA has been extracted.
In the context of a blood DNA extraction, a washing buffer might be used to wash away red blood cells, proteins, and other impurities that may still be present after the DNA has been isolated. Washing buffer is also used before elution to purify the DNA further.
Tris-EDTA (TE) Buffer
Tris : buffer that maintains a stable pH (typically around 8.0), which is important for preserving the integrity of DNA.
EDTA : A chelating agent that binds divalent cations (like Mg²⁺ and Ca²⁺), which are necessary for nucleases to degrade DNA. By sequestering these ions, EDTA helps protect the DNA from degradation.
TE buffer is used to resuspend, store, and protect DNA after extraction. It is not intended to remove contaminants, but to stabilize and preserve DNA.
The fact that your DNA samples appear creamy and the bands on gel electrophoresis are unclear could be due to several factors related to contamination or improper sample processing :
- Incomplete Lysis or DNA Precipitation
- Use of TE Buffer as a Wash
- Improper DNA Elution
- Overloading of the agarose gel
Look at each of these different steps to optimize your blood DNA extraction
Tris EDTA is used to prevent the degradation of DNA by inhibiting nuclease enzyme also used for storage purpose while washing buffer is added to remove the impurities and the contaminants.
In blood DNA extraction, Tris-EDTA (TE buffer) is primarily used for storing DNA due to its ability to maintain pH and protect DNA from degradation. However, it is not ideal for washing because it may not effectively remove proteins and contaminants, leading to unclear, creamy DNA samples and poor band clarity in gel electrophoresis. In contrast, a washing buffer is specifically formulated to remove impurities, often containing salts and detergents that help produce clearer DNA samples.
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