I suppose you might need to look for answers yourself by reading some among hundreds of article online. The question enlists so many points of basic understanding and indicate the lack of fundamental understanding of techniques. Please go though the literature on primarily the manufacturers website to get the details of the instrument and principles.

Abhijeet Singh Thank you for trying to answer my question. Perhaps I was not specific enough in my question and you did not understand what I mean.

I have already read the literature at Thermofisher´s webpage.

The Qubit measures fluorescence and is supposed to be more sensitive because the samples are prepped beforehand by adding fluorescent dye and buffer which allows the Qubit to measure only the sample binding to the dye unlike a spectrophotometer ( like the Nanodrop) that would measure anything that reflects UV at, say a 260 nm wavelength for nucleotides.

However, in the case of DNA and RNA the Nanodrop would also measure free nucleotides or phenols (maybe) which can make the concentration of the sample higher than it actually is.

I have also attempted to differentiate between the Nanodrop and Qubit by measuring a series of dilutions of BSA supplied by Thermofisher with the concentrations already told by the company. And I did not see a major difference in sensitivity between the two instruments except for at lower concentrations, where the Qubit can still detect protein while the Nanodrop does not.

I am going to continue reading articles. But I wanted to know the personal experiences of a wide array of researchers who use the Qubit and Nanodrop in their own labs but did not publish anything to get an idea besides the articles.

Ahmed Ibrahim Akhtar

Thanks for the clarification/question buildup and I appreciate that. Now, surely this question looks like from someone who made their efforts in getting answers and not directly asking question on RG.

You have got everything right in your understanding and explanation. Regarding the standards you mention, I can share my understanding on the measurement ability of both methods.

Nanodrop is not a bad method. It has been used for a long time and still widely in use. For standards, we don't see any difference because in these solutions are very pure with only nucleic acids as absorbent. However, for samples, there can be other unknown chemicals which can cause differences in the respective assays. In case of pure samples, I think both methods are still comparable. additionally, even same sample can give variable results in both assays if treated differently. Thus, such differences are dependent of sample type, content/quality of extracted NAs, pipetting etc. mainly the user related parameters. I guess therefore such things are not mentioned in articles and are experienced based.

Abhijeet Singh Thank you for the answer.

I think, according to my own time using the Qubit and Nanodrop that chances of human error are perhaps lower with the Qubit because the instrument refuses to progress if even one of your eight tubes containing standards is incorrect. It´s annoying, sure but probably less error-prone.

Thanks also for asking me to elaborate. I appreciate learning how to ask questions better.