I am usually working with zebrafish DNA and qiagen minipreps often yield 500 ng/ µl DNA. I use 5 ml of LB incubated overnight. Since I started doing minipreps with Salmon DNA, using the exact same protocol and vectors, the yield has decreased to around 50 ng/ µl or lower. I tried prepping different samples and multiple times but still no result. Does anyone have a idea how I can solve this?
Dear Matthijs
if you are using same vector is not easy to undertand why this difference happens. Do you know if there is a huge differences in terms of gc% beetween the 2 genomes that can explain lower dna stability?
i can just suggest to you:
to use a different e.coli strain per propagation (you can try hb101, sure , scarab express) to see if your plasmid is more stable in this strains.
to try the omega biotek ezna mini kit2 which alllw you to start from 10ml of lb o/n and generally in my experience over perform all the other mini kit.
good luck
Manuele
Hi Matthijs,
If you do minipreps with both set of vector (with zebrafish and salmon DNA) would you get high yield in one and not the other? or would you get low yield with both? If the later is the case, then it is independent of the vectors and you should check the antibiotics you use to select the plasmids. They could be outdated or spoiled and that would explain you have less plasmid-containing bacteria and therefore less DNA yield. Alternatively, you can perform a homemade miniprep (cloroform extraction) and check the yield there. This would help to rule put that the miniprep column from the kit is not retaining efficiently he DNA.
Cheers,
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