I use a kit for extraction of cf DNA by Qiagen using magnetic beads. For an elution of 40 microliters concentration and absorbance ratio was very poor for the 10 samples extracted.
I need to know if anybody has used the same kit and their review on the same.
Hi Sonam Kille,
I didn't use the Qiagen kit. However, I have tested the MagMAX (ThermoFisher) kit for cfDNA isolation using magnetic beads. How many DNA nanograms you obtained from your protocol? Indeed, the cfDNA concentration from plasma is very low (~5-10 ng/uL). Which was your cfDNA source? It also depends on the storage conditions of your sample. For instance, did you work with fresh or frozen samples?
We can talk more about that.
Hi Alexis Murillo Carrasco,
The concentration is indeed around 5-10ng. So the concentration isn't bad I guess. I have worked on frozen samples.
I was able to use the cfDNA for downstream application like ddPCR. I got expected results.
thank you
Hi Sonam,
Yeah, probably that's right. What is your input sample volume?
I haven't used the Qiagen kit, but the Apostle cf kit (https://www.beckman.com/reagents/genomic/dna-isolation/from-plasma/performance) which uses magnetic beads. I was starting from 500ul plasma and getting very low amounts - around 1-2ng/ul as measured by Quibit but this was enough to then bisulphite treat and PCR. You won't get much cf DNA from plasma. Your starting material is important, we were collecting blood in streck tubes to try maximise cf DNA. https://www.streck.com/products/stabilization/cell-free-dna-bct/
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