I am planning to extract DNA with QIAamp Mini DNA extraction kit (Qiagen) from DNA Shield microbiome preserved samples. My samples contain small amount of cell, so my goal is to reduce the loss of cells by extracting DNA from the maximum amount of sample possible. QIAamp Mini kit recommend using a maximum of 200ul of liquid sample for the extraction, but I contacted the company and they told me I can increase it until 400ul if keep the proportions by doubling the volume of the kit reagents and pass all the liquid through the membrane in two centrifugation steps.
I aim to minimize both the amount of liquid and beads to be able to use the maximum amount of sample during DNA extraction and reduce the loss of DNA from my low concentrated samples. So, I would like to perform bead-beating prior DNA extraction in the minimum about of liquid and beads.
Do you know what is the minimum amount of liquid and beads (0.1 + 0.5mm) I can use for bead-beating in a 2ml vial?
Good question, and I am not aware of a clear answer. Bead beating procedures have been optimized for duration (Article Optimisation of a bead-beating procedure for simultaneous ex...
), bead size (Article Optimizing an enclosed bead beating extraction method for mi...
) but I dont know of any studies optimizing bead cc., in part because the results would heavily depend on the starting material (eg feces vs water vs soil will give you very different bead beating requirements). My only suggestion would be to cut bead cc to half but increase duration, and see if that gets you the result you hope to achieve.
Dear Péter Gyarmati
Thank you for yor answer.
After reading several sources and discussing with colleges, I got the conclusion that a good liquid-bead proportion could be around 60% of the volumen of liquid used, which match quite well with your suggestion of cutting bead cc to half.
Not talking about proportion of liquid and beads, but total final volume. Do you know if is there any minimum amount of liquid can be used for bead beating in a 2ml vial? For example, would 400ul of liquid and 200ul of beads (total volume 600ul) be too little volume to lyse the cells?
Thank you in advance.
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