Hello, I am currently performing DNA extraction from an anaerobic digester using the FastDNA spin kit for soil, which involves 2 SEWS-M wash steps. Prior to extraction, I subjected the samples to three washes with a 0.85% KCl solution. However, when assessing the purity of my extracted DNA with a Nanodrop, I observed a favorable 260/280 ratio of 1.8 but encountered issues with a very low 260/230 ratio of 0.5 and a high A230 value of 3.
I am seeking advice on how to improve the 260/230 ratio. Should I consider adding a third SEWS-M wash step, switching to a phosphate buffer pretreatment instead of 0.85% KCl, or potentially combining both methods for sample pretreatment? I've also experimented with a 5.5 M Guanidine Thiocyanate treatment, but this didn't yield any improvement. In fact, after this treatment, the A230 and 260/230 values remained the same, and the 260/280 ratio decreased from 1.8 to 1.7. Any suggestions or guidance would be greatly appreciated. Thank you.
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