Why orcinol method is used for RNA estimation?
This is a general reaction for pentose and depends on the formation of furfural. When pentose is heated with concentration HCl, orcinol reacts in presence of furfural in presence of ferric chloride as a catalyst purine to produce green color only the purine nucleotide.
What is orcinol method?
Principle: The method usually involves the pentose conversion and ribose in the presence of hot acid to furfural that again reacts with orcinol to form a green colour. The colour that is formed largely depends on HCL concentration, ferric chloride, orcinol and the time of heating at 100oC to some extent.
How do you quantify RNA concentration?
The traditional method for assessing RNA concentration and purity is UV spectroscopy. The absorbance of a diluted RNA sample is measured at 260 and 280 nm. The nucleic acid concentration is calculated using the Beer-Lambert law, which predicts a linear change in absorbance with concentration (Figure 1).
Which method is used for estimation of RNA?
Ultraviolet (UV) absorbance can be used to measure DNA, RNA or protein concentration. For nucleic acids, the three main wavelengths of interest are 260nm, 280nm and 230nm. Absorbance at 260nm is used to measure the amount of nucleic acid present in the sample.
How is the estimation of RNA by orcinol method?
But one of the widely used reactions is those of RNA with Orcinol. So, the basic objective of this test is to estimate RNA by orcinol method. Principle: The method usually involves the pentose conversion and ribose in the presence of hot acid to furfural that again reacts with orcinol to form a green colour.
How is the green colour of orcinol determined?
Principle: This is a general reaction for pentoses and depends on the formation of furfural when the pentose is heated with concentrated hydrochloric acid. Orcinol reacts with the furfural in the presence of ferric chloride as a catalyst to give a green colour, which can be measured at 665 nm.
How much RNA can be diluted in one ml?
Prepare a series of serially diluted RNA standards, but with a range from 1.0 mg/mL down to 0.125 mg/mL. Prepare a serial dilution of your sample RNA.
Which is the most common method of RNA measurement?
The most frequent approach for RNA measurement will be discussed in this post. This is a general pentose reaction that relies on the production of furfural when the pentose is cooked in strong hydrochloric acid.