I've been doing a side by side analysis of Chl content of rice leaves using traditional UV-VIS spectrophotometer and Nanodrop, and so far the values that I'm getting are far from each other. The samples are diluted. Could this be the cause of the discrepancy in values?
In case of nano drop we generally go for DNA, RNA, Protein readings as after isolation of these macromolecules we invest them little to know their concentration and rest can be used for further experiments we have planned. For analysis of chl you may safely go for UV-Vis spectrophotometer. After getting concentration readings from UV-Vis spectro if you dilute the sample and take reading from nano drop it will obviously give different readings from UV-Vis Spectro readings. Therefore, I advise you for Chl readings just have data from UV-Vis spectrophotometer and rely on it. Go for at least three replicates and calculate standard deviation as well.
Concentration readings from UV-VIS spectro. Dilute samples. Take reading from Nano Drop.
I have used NanoDrop for concentration determination of protein samples.
If you have enough sample, I would go for UV/Vis for a more accurate reading.
If you want to rely on NanoDrop, you will obviously have to correct for your dilution. I would take at least three repetitions and average. Also errors can occur if your sample is not evenly distributed, so it could be advisable to take a small amount from the top, bottom and center of your tube in order to exclude this source of error.
Thank you everyone. Based from what i've read, my results, and from your answers I think I would rather use the Uv-Vis to get a more accurate and precise result. Thank you!
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