Hi, I would suggest using a fluorescent based method to measure the concentration of your DNA. It is much more accurate. If your DNA extract is clean and has high concentrations, then you can use Nanodrop. If not, definitely use Qubit. I use the Qubit dsDNA HS Assay Kit.

Dear Ahamed,

The answer to your questions is Yes. You have to make DNA with minimum contaminations of minerals or other chemicals used for purification. Residual chemical contamination from nucleic acid extraction procedures may result in an overestimation of the nucleic acid concentration and/or negatively influence downstream analysis.

The purity ratios are important indicators of sample quality. The ratio of 260-280 nm ~1.8 is generally accepted as “pure” for DNA; a ratio of ~2.0 is generally accepted as “pure” for RNA. If the ratio is appreciably lower in either case, it may indicate the presence of protein, phenol, or other contaminants that absorb the waves at 280 nm.

I would like to suggest find the your actual problems while DNA purification then fixes the problem before the shift to other quantification methods. It gives more precise results of your downstream analysis.

Hello

I remember a long time ago, when there was not nonodrop or qubit machines, we have successful results for most PCR reactions. The only reliable tool for integrity-quantity assessments was comparative electrophoresis. Jennifer Tu is right but for a routine PCR, qubit would be expensive. Since we have an Epoch (similar mechanism to a nanodrop) instrument in our lab, there was no problem so far.

Thank you all @ Jennifer Tu, Shan Lakmal Edirisinghe , Ali Javadmanesh

Hi Ahmed Hasan Mohammed,

DNA needs to be in a good concentration and quality for PCR. You can use nanodrop, qubit or other system. Another important thing is to storage the DNA at -20 or -30 when you are not using it. If you are using the DNA try to keep at 4C to preserve it. Try to reduce the changes of temperature because it will affect the concentration.