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How to determine the purity and yield of DNA?

5 years ago in Case Study By Heena


Is it possible to determine the purity of a DNA sample? Also, which is the best technique that can be used to identify the yield of the DNA sample.

Yes, you can definitely measure the purity of the DNA sample. There is a technique known as absorbance which is widely used to determine the purity of DNA sample. 
This is one of the simplest methods. It includes equipment such as UV transparent, UV lamp and purified DNA solution. 
All you have to do is measure the absorbance from approximately 230 nm to 330nm. Then take the ratio of absorbance reading. 
Next, calculate the DNA purity by using the formula,
DNA purity = (A1reading – A2 reading) / (A3 reading – A2 reading)
 

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By Priya Answered 5 years ago

Yes, you can definitely measure the purity of the DNA sample. There is a technique known as absorbance which is widely used to determine the purity of the DNA sample.  This is one of the simplest methods. It includes equipment such as UV transparent, UV lamp and purified DNA solution.  All you have to do is measure the absorbance from approximately 230 nm to 330nm. Then take the ratio of absorbance reading.  Next, calculate the DNA purity by using the formula, DNA purity = (A1reading – A2 reading) / (A3 reading – A2 reading)  


By Bindya Answered 5 years ago

You can use agarose gel electrophoresis to determine the yield as it is considered as the best method used for the same.  But I would suggest you use the absorbance method as it is easy to use and requires simple equipment.


By Kunal Answered 5 years ago

The best method to determine the DNA yield is agarose gel electrophoresis. Here, the gel electrophoresis connected with external power supply,  running buffer and analytical-grade agarose. The sample is loaded in the agarose gel and exposed to electric gel.  The negatively charged DNA migrates towards the anode. Small fragments of DNA migrate faster and RNA, nucleotides migrate at different rates.  The yield can be determined after the completion of gel electrophoresis


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