Hello everybody. I'm doing DNA extractions from rizobacteria. The problem is the 260/230 ratio. I have both low and high ratios (eg. 0.76 or 3-4.09). When I search information in Thermo Scientific it said that values above 2 in this ratio doesn't mean that you will have problems but it seems the opossite. When I tried 16S-PCR I didn't have any band. I will appreciate some recomendations for both cases to clean my DNA (that seems the only solution).
About the samples, I always clean them before extraction with saline solution (three times).
Thank you for your time!
Have a nice day!
Hi it is not clear which kit or procedure you have used for DNA extraction.
You can try alcohol precipitation if it was not used before.
Good morning Dr. Marcela. I used the Purelink Genomic DNA mini kit (Invitrogen). About the alternative you mentioned it, I haven't done it yet. I'm going to try that. Thank you very much.
Best regards.
You can repeat Tris saturated phenol -chloroform extraction step once again and try precipitating it with alcohol to get rid of RNA and Protein(for RNA extraction use acidic phenol) if you want to selectively isolate RNA use lithium chloride or use RNase to remove it in final step.
You may be having a high load of impurities in your final DNA elute which contributes to the fast degrading of your DNA, you might want to change the extraction kit or perhaps even try colony PCR since that will give you a high yield of DNA fo your PCR experiments.
As per the recommendation of Venkateswa rarao Nuthakki Repeat the Phenol- chloroform step then precipitate by using Sodium accetate and Ethanol. During the purification process please make sure that there should not be any traces of phenol.....
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