I am doing DNA extration from whole saliva using Vivantis DNA extraction kit for Viral nucleic acid purification. After DNA extratcion on PICO drop the values of DNA is 333 microgram per microliter or 423 microgram per microliter, whereas the ratio of A260/A280 is 3.3 or above 3 and ratio of A260/A230 is above 3. Is these values are normal? or high ratios mean?
Is this high ratio of A260/A280 and A260/230 is okay for future PCR analysis?
Dear Sadia
A260/A280 and A260/230 ratio higher that 2 are very good.
Higher ratio means higher sample purity.
in the following link
http://www.nanodrop.com/Library/T009-NanoDrop%201000-&-NanoDrop%208000-Nucleic-Acid-Purity-Ratios.pdf
you can find some basic information.
best regards
manuele
Thank you Manuele for your reply but someone guide me that this much high ratios in both is not good for PCR.
Sample 1: DNA (307) A60/A280 (4.7) A260/A230 (4.88)
Sample 2: DNA (483) A60/A280 (3.8) A260/A230 (4.2)
Sample 3: DNA (727) A60/A280 (3.3) A260/A230 (3.5)
Hi Sadia: Here is some useful papers
http://www.nanodrop.com/Library/T009-NanoDrop%201000-&-NanoDrop%208000-Nucleic-Acid-Purity-Ratios.pdf
http://www.nanodrop.com/Library/T042-NanoDrop-Spectrophotometers-Nucleic-Acid-Purity-Ratios.pdf
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Hi Sadia
Some papers in attachment 1
http://www.nanodrop.com/Library/T042-NanoDrop-Spectrophotometers-Nucleic-Acid-Purity-Ratios.pdf
2
http://www.nanodrop.com/Library/T009-NanoDrop%201000-&-NanoDrop%208000-Nucleic-Acid-Purity-Ratios.pdf
4
https://www.ogt.com/resources/literature/483_understanding_and_measuring_variations_in_dna_sample_quality
5
http://www.biotek.com/resources/application-notes/nucleic-acid-purity-assessment-using-a260/a280-ratios/
6
https://classes.soe.ucsc.edu/bme220l/Spring11/Reading/PowerWave200_Nucleic_Acid_Purity_Assessment.pdf
7
https://www.biotechniques.com/multimedia/archive/00010/96201bm06_10160a.pdf
8
http://molecularbiology.forums.biotechniques.com/viewtopic.php?t=19137
9
http://www.gelifesciences.com/file_source/GELS/Service%20and%20Support/Documents%20and%20Downloads/Handbooks/pdfs/Spectrophotometry.pdf
The 260/230 ratio is used to indicate the presence of unwanted organic compounds such as Trizol, phenol, Guanidine HCL and guanidine thiocyanate. Generally acceptable 260/230 ratios are in the range of 2.0 – 2.2.
In buffered solutions, pure dsDNA has an A260/ A280 of 1.85–1.88 and pure RNA has a ratio of around 2.1.
In my opinion I would suggest further purification of the DNA samples before going to PCR to avoid further complications.
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