Does autoclaving destroy RNA?

Autoclaving does NOT fully destroy nucleic acids: PCR analysis demonstrates that even after autoclaving, larger DNA fragments can be identified, especially when nucleic acids are protected by protein envelopes (e.g. viruses) or within microorganism cell walls (e.g. bacteria).

How does orcinol helps in the estimation of RNA?

HiPer® RNA Estimation Teaching Kit is designed for rapid and accurate determination of RNA by orcinol reagent. This method depends on the conversion of pentose in the presence of hot acid to furfural which then reacts with orcinol to yield a green colour. The colour intensity can be measured at 665 nm.

Is RNA positive in Benedict’s test?

Benedict’s – Positive = free ribose, fructose, glucose; Negative = RNA, DNA, ATP, etc.

Does RNA react with diphenylamine?

RNA has been demonstrated to react with diphenylamine when acid hydrolysis is performed for 1 hour or more at 100°C. This reaction can be used for quantitative analysis of RNA, since there is a linear relationship between RNA concentration and absorbance.

Can you vortex RNA?

 Do not vortex Trizol lysates or RNA samples to avoid shearing.  After extraction, keep RNA samples on ice at all times.

How can RNA denature?

b. Heat denature samples at 65-70°C for 5-15 min. Denaturation for 5 min is typically sufficient for simply assessing RNA on a gel, but a 15 min denaturation is recommended when running RNA for a Northern blot. The longer incubation may be necessary to completely denature the RNA.

How do you prepare orcinol reagent for RNA?

In another test tube add 3 ml of water to keep it as blank. Now take 6 ml of orcinol acid reagent and add to each tube. Again add 0.4 ml of 6% alcoholic orcinol to every tube. Now shake every tube so that everything is mixed up very perfectly and keep it in boiling water bath for about 20 min.

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.

What are the chemical test used to identify the presence of DNA and RNA?

The (Dische) Diphenylamine Test is used for determining the presence of nucleic acids. The presence of DNA will turn a clear solution blue. The more DNA present the darker the color. Another nucleic acid, RNA, will turn green.

Can diphenylamine reaction be used to distinguish RNA from DNA?

Dische Diphenylamine Test For DNA Acidic conditions convert deoxyribose to a molecule that binds with diphenylamine to form a blue complex. The intensity of the blue color is proportional to the concentration of DNA. The Dische’s Test will detect the deoxyribose of DNA and will not interact with the ribose in RNA.

How does diphenylamine react with DNA?

The deoxyribose in DNA in the presence of acid forms β-hydroxylevulinaldehyde which reacts with diphenylamine to give a blue colour with a sharp absorption maximum at 595nm. In DNA, only the deoxyribose of the purine nucleotides react, so that the value obtained represents half of the total deoxyribose present.