BACKGROUND
The concentration of RNA present is directly proportional to the absorbance of light carried out in a spectrophotometer. This Beer Lambert’s Law. The basic objective of this test is to estimate the RNA by using UV Spectrophotometer.
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REQUIREMENTS
RNA sample
TE buffer
UV spectrophotometer
PROCEDURE
At first take out 10 microliter of total RNA and then dilute it with 990 microliters of TE buffer which contains 10 mM Tris, 1 mM EDTA at a pH of 7.0. In centrifuge tube fill it and then put it in spectrophotometer for measuring the absorbance at 260 and 280 nm. For calculating the total RNA use the following formula.1
Total RNA (ug) = (A260) (40 µg) (100) (0.05 ml).
Concentration= total ug/50 µl or mg/ml.
CONCLUSION
With this method we can easily estimate RNA.
REFERENCES
- Wilfinger WW, Mackey K, and Chomczynski P (1997) Effect of pH and ionic strength on the spectrophotometric assessment of nucleic acid purity. Biotechniques 22:474 481.
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