Team:Oxford/protocols/Nanodrop: Finding DNA Concentration
From 2014.igem.org
Nanodrop: Finding DNA Concentration
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- Ensure the program is selected to read “NUCLEIC ACID” where it says “Sample Type” (Top right of screen)
- Load 2 µl of your blank onto the raised pedestal. (NOTE: this blank should be the solvent your DNA/RNA is suspended in. For example this could be EB buffer if you have just done a MiniPrep)
- Lower the metal lever until it gently rests on the sample pedestal and click ‘BLANK’ to zero the spectrophotometer reading.
- Using a tissue blot off the liquid from the pedestal and the contact on the underside of the lever.
- Now load 2µl of your nucleic acid sample in the same manner as the blank and close the lever
- Click ‘MEASURE’. After a short while it will display a concentration in ng/µl in the bottom right hand corner.
- Also displayed is the UV/Vis spectrum. One should expect a clean ‘bell’-like curve that indicated a clean sample. If a rough and wavy spectrum is seen the sample is likely contaminated.
- Blot off the sample from the contacts as before and repeat for other samples.
- At the end of your use wash the contacts by adding 2µl water to the contacts and closing the lever a few times before blotting off the water to leave the machine dry.