I am using CTAB method for RNA extraction. However, during RNA extraction DNA also isolated and DNase treatment lowers the quantity of RNA. Kindly let me know the reason why DNA is coming along and at which step in extraction I should be more careful.
Read the following article and compare it with your method.
Article Methods of RNA Purification. All Ways (Should) Lead to Rome
What is the pH of your extraction buffer? I suspect it is pH 7 or close to it. Raise the pH to 8 or even 9 and use isopropanol, not ethanol. And please report your results back.
You need to check your Reagents and PH of the buffer solution (be sure to all buffer and reagents are fresh). Also, the problem can sometimes be avoided by sterile conditions.
Generally, some methods used to reduce or remove DNA contamination including: DNAse I digestion, gel fractionation, phenol: chloroform extraction, and lithium chloride precipitation.
The use of lithium chloride in RNA precipitation is recommended.
Lithium chloride is a selective precipitant of RNA with DNA being left in the supernatant. A major advantage of lithium chloride is that it does not efficiently precipitate either protein or DNA.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA