Hello!
I'm using Ultra-Deep Microbiome Prep kit for the isolation of DNA from mice skin biopsies. I was doing it yesterday for the first time and I didn't achieve to any DNA yields. So I would like to ask you.. Is here someone who has experiences with this kit? I isolate the DNA from frozen samples, is it big problem? And maybe I don't understand one step in the protocol. Step 10: "Pipette lysate into a Spin column. Pipette the fluid phase in the column. Avoid transfer of any unresolved particles." Well, when I was doing it, I had just a homogeneous solution without any particles, so I transfer it all to the column. Was it right?
Thank you very much!
Isolation of good quality DNA/RNA is extremely important for performing trouble-free downstream processes. The isolation of DNA using a customized kit is not a big deal for researchers. However, it requires care at multiple steps.
- Make sure you are not using too little or too high amount of the tissue/bacterial cells than recommended in the protocol.
- You can prolong incubation time in the lysis step.
- If you are precipitating DNA using ethanol, it is highly recommended to use molecular biology grade ethanol.
- The ratio of ethanol/isopropanol used in the DNA precipitation step can be increased.
- At elution, the incubation time after adding the elution buffer on the column can be increased.
And 'Yes', at step 10, you are doing according the protocol.
P.S: You can practice on some samples other than your actual experiment until you optimize you protocol.
Good Luck
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