I want to demonstrate a transcriptional regulation by the coinfiltration with Agrobacterium in N.benthamiana the TF and a promotor sequence. But I don't know which strain of Agro is better for that GV3101 or 3010?
GV3101 worked well in my several projects (N. tabacum). Have not used 3010.
Thank you very much Dr Yuan-Yeu Yau, for your comment. It really is very helpful to me
We use GV3101 routinely for Nicotiana infiltrations, works great. We haven't used GV3010, so I can't comment on that. A few tips for infiltrations. Make sure you're working from plates freshly streaked from glycerol stocks (which is best practice!). I had a bad experience when re-streaking a plate from and older one - they grow fine, with all the selective antibiotics (we use Spec100 for binary vector and Gentamycin 20mg/L for the helper plasmid). But when inducing the cultures with acetosyringone prior to infiltration, these strains failed to produce the normal funky smelling volatiles. And transformation was really poor. Basically they failed to become properly infectious. I did a side by side with a GFP construct too - the fresh stock produced the volatiles and GFP was super strong, but the other required very high magnification to see any GFP at all. Other colleagues have had similar anecdotal issues when re-streaking Agrobacterium cultures (I do not think this is unique to GV3101). I'm now very particular when establishing new strains and when using them from the freezer. The other tip, Agro grows very well on Agar plates. We just use a confluent loop of cells scraped off a plate and resuspend in 10mM MgCl2 with 200 uM Acetosyringone. Much easier than liquid cultures, pelleting cells etc.
Thank you very much Dr Nick W Albert for clarifying my doubts and for the excellent tips. I will take them all into account.
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