What cause the Gloo after incubation using Qiagen DNA extraction kit?

28 June 2023 1 10K Report

I was performing the DNA extraction of cells with a concentration of 5x10^6. After adding AL buffer and proteinase K an incubation was done. After the incubation period at 56 degree Celsius, I added ethanol and did a pipette mix and realised there was a huge gloo and it can't pass through the flow through tube after centrifugation. May I know if something was done wrongly that cause the gloo formation?

Paul Rutland

I think that this is caused by protein precipitating on the dna and forming an insoluble mix. You may need to use a smaller number of cells or allow longer at the proreinase K degradation stage, Possibly also you may have a problem with insufficient SDS containing buffer because protK works best on partially degraded samples

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