Hi every one,
I am having a tough time with my DNA extraction.
I am extracting DNA from stained cytological slides using QIAmp Mini Kit. I am performing RNAse treatment.
Well, what is happening is that I am having odd results from my Nanodrop measurment!
Apart from low concentration in some of the samples, sometimes I have got very high 260/280 ratio values (even 7!) or very low values for 260/230 ratio.
Here it is an example:
Concentration (ng/ul )71.48
A260 1.430
A280 0.251
260/280 5.70
260/230 0.14
I am using different protocols, and I am beeing very careful during all the procedure but my results are not changing. I've read a lot about low ratio values and contamination, but i really don't know how to interpret tha high values (especially that high). My doubt is that my Nanodrop is having some issues.
Am I right? Or there is someting that I am missing?
Thanks for any help.
You may run your sample in a gel to verify the quality. If you are following your protocol properly and able to see a good electrophoresis band, you may proceed for downstream actions using your DNA and check out if your Nanodrop needs to be fixed or not. Best wishes.
hi
try to measure your samples in another Nanodrop ..your current one seems to need calibration!
2 things about nanodrops. Often after much use the quartz optical window accumulates protein and dna from the samples. You should try cleaning it with HCl .
Secondly they are very sensitive to zeroing. it is essential to zero the machine just before sample measurement and with exactly the same solvent that the dna is dissolved in. You cannot zero with water and measure dna dissolved in TE for instance. If the dna is dissolved in elution buffer from a kit then zero with elution buffer
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