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Quantitative Estimation of DNA and RNA

  • Aakanchha Jain
  • Richa Jain
  • Sourabh Jain
Protocol
  • 336 Downloads
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

The purine (adenine and guanine) and pyrimidine (cytosine, thymine, and uracil) bases of DNA and RNA can be estimated quantitatively and qualitatively using UV spectrophotometer, because the conjugated double bonds of nucleic acids tend to absorb ultraviolet light between 220 nm and 300 nm. On solubilizing the DNA in citrate buffer and measuring its optical density at 260:280 nm, the ratio of 1.85 indicates that the isolated DNA is pure. The ratio changes to ~2.0 for RNA and decreases below 1.85 for protein contaminations if any. Nomogram is the scale which indicates the amount of both RNA and DNA in a sample at the same time.

Principle

The purine (adenine and guanine) and pyrimidine (cytosine, thymine, and uracil) bases of DNA and RNA can be estimated quantitatively and qualitatively using UV spectrophotometer, because the conjugated double bonds of nucleic acids tend to absorb ultraviolet light between 220 nm and 300 nm. On solubilizing the DNA in citrate buffer and measuring its optical density at 260:280 nm, the ratio of 1.85 indicates that the isolated DNA is pure. The ratio changes to ~2.0 for RNA and decreases below 1.85 for protein contaminations if any. Nomogram is the scale which indicates the amount of both RNA and DNA in a sample at the same time.

Combined Viva Voce for Experiment Determination of Melting Temperature of DNA and Quantitative Estimation of DNA and RNA

  1. Q1.

    What is meant by melting of DNA?

     
  2. Q2.

    What is the principle of determining melting temperature of DNA?

     
  3. Q3.

    Which buffer is used for dissolving DNA?

     
  4. Q4.

    What is the most abundant form of DNA in nature?

     
  5. Q5.

    _____________temperature range is used for determining melting temperature of DNA.

     
  6. Q6.

    What is the absorbance ratio of pure DNA at 260:280 nm?

     
  7. Q7.

    The value of 2.0 obtained for 260:280 ratio at UV is for_____________

     
  8. Q8.

    The value less than 1.85 indicates_____________contamination in isolated DNA sample.

     
  9. Q9.

    The scale used for estimation of RNA and DNA in a sample at a time is called as_____________

     
  10. Q10.

    Which wavelength is used for UV estimation of DNA and RNA?

     

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Aakanchha Jain
    • 1
  • Richa Jain
    • 2
  • Sourabh Jain
    • 3
  1. 1.Bhagyoday Tirth Pharmacy CollegeSagarIndia
  2. 2.Centre for Scientific Research and DevelopmentPeople’s UniversityBhopalIndia
  3. 3.Sagar Institute of Pharmaceutical SciencesSagarIndia

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