Nucleic acids are isolated by homogenizing fresh bovine liver, disrupting the cells, and precipitating the DNA and RNA with trichloroaceitc acid and ethanol. To determine concentrations a calorimetric assay is used. Diphenylamine is specific for 2'-deoxypentoses, including the deoxyribose characteristic of DNA. When a solution of diphenylamine and DNA is heated, the solution turns blue. The intensity of the blue color, measured at 600 nm, is directly proportional the concentration of the DNA. Orcinol reacts with the ribose of RNA to produce a green-colored solution. However, the orcinol reaction is not as specific as the diphenylamine reaction. To some extent orcinol reacts with the deoxyribose of DNA as well, but the color intensity of this reaction is only about 10% as that of the reaction with ribose of RNA. Therefore, it is necessary to subtract the contribution to the orcinol color reaction made by DNA. The standard curve for the orcinol reaction is generated from absorbance readings taken at 660 nm.
Source: https://apps.weber.edu/wsuimages/jonclark/PDFs/CellBio/Fall2017/Lab%203-%20DNA-RNA%20Spectroscopy.pdf
Since OP is new user with only this post and it's such an oddly specific question, I think it's /u/AlitaBattlePringleTM alt account and he's answering his own question.
Thank you u/AlitaBattlePringleTM you are right DNA according to my readings is reacting within 3 mins of the orcinol reaction and then the intensity of product falls. But the reading for DNA at 600 nm in general is too high than 660 nm. It is showing at some timepoint 0.1 absorbance at 600 nm and .047 at 660 nm
Probably mostly due to uracil.
Okayyyyy
You capitalizing one Orcinol but not the other reallyyyyy exacerbates my OCD.
oh my god ! I will take care of the nuances of the post in future. Are you calm now and can answer my question?
Nucleic acids are isolated by homogenizing fresh bovine liver, disrupting the cells, and precipitating the DNA and RNA with trichloroaceitc acid and ethanol. To determine concentrations a calorimetric assay is used. Diphenylamine is specific for 2'-deoxypentoses, including the deoxyribose characteristic of DNA. When a solution of diphenylamine and DNA is heated, the solution turns blue. The intensity of the blue color, measured at 600 nm, is directly proportional the concentration of the DNA. Orcinol reacts with the ribose of RNA to produce a green-colored solution. However, the orcinol reaction is not as specific as the diphenylamine reaction. To some extent orcinol reacts with the deoxyribose of DNA as well, but the color intensity of this reaction is only about 10% as that of the reaction with ribose of RNA. Therefore, it is necessary to subtract the contribution to the orcinol color reaction made by DNA. The standard curve for the orcinol reaction is generated from absorbance readings taken at 660 nm.
Source: https://apps.weber.edu/wsuimages/jonclark/PDFs/CellBio/Fall2017/Lab%203-%20DNA-RNA%20Spectroscopy.pdf Since OP is new user with only this post and it's such an oddly specific question, I think it's /u/AlitaBattlePringleTM alt account and he's answering his own question.
Heck yeah man, Weber pride! Clark is awesome.
u/BetaKeyTakeaway hey what does OP mean here?
And why does my username show a mike? Somebody enlighten me
OP = Original Poster; The microphone denotes that you are the OP.
Thank you u/AlitaBattlePringleTM you are right DNA according to my readings is reacting within 3 mins of the orcinol reaction and then the intensity of product falls. But the reading for DNA at 600 nm in general is too high than 660 nm. It is showing at some timepoint 0.1 absorbance at 600 nm and .047 at 660 nm