Hello Jyoti
DNA extraction involves three basic steps:
1. Lysis in which the cell and the nucleus are broken to release the DNA.
2. Separation of DNA from proteins and cellular debris.
3. Precipitation of DNA using alcohol.
You can use the traditional Phenol-Chloroform method for DNA extraction from tissue. The yield of DNA is high and the quality is good. The reagents that are required for DNA extraction are simple and not very expensive. They are usually found in the laboratory which include Phenol, Chloroform, Isoamyl alcohol, ethanol, sodium acetate and potassium acetate.
Please note you need to carry out proper lysis of the tissue to obtain good results.
Thank you.
Hello Jyoti Singh. How are you doing today? We use an adapted protocol from the following paper. https://pubmed.ncbi.nlm.nih.gov/9358185/
This method uses a saline buffer, SDS, Proteinase K, Isopropanol, and ethanol.
Alternatively, you can use a 50mM of NaOH solution to extract DNA. Incubate the tissue in the 360ul of NaOH solution at 95°C for 5 minutes. Add 20ul of Tris-HCl pH 8,0 to neutralize the reaction. Centrifuge at 12000 RPM for 5 minutes and the DNA will be suspended in the solution. You can transfer the supernatant to other tube, do not touch the organic phase in the bottom. Do not use much tissue to extract DNA, just a small fragment will be enough, and make sure the tissue will be degraded. It will appear as a jelly.
Look in this publication. May be used on tissue as well and does not require phenol_chloroform.
Miller SA, Dykes DD, Polesky HF (1988) A simple salting out
procedure for extracting DNA from human nucleated cells.
Nucleic Acids Res 16:1215
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