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Gordonia DNA not cutting

| posted 19 Nov, 2020 16:12
Anyone have any trouble getting a good RE digest of Gordonia DNA? I've got good DNA from M. foliorum, and we're trying Gordonia rubripertincta for the first time this year. I've noticed it's a bit waxier than M. foliorum. I'm getting OK DNA concentrations with a new Wizard kit, but my Mf DNA cuts where my Gr does not. Any tips/tricks I'm missing?
Thanks!
| posted 19 Nov, 2020 16:58
Hi Amy,

Would you mind sharing a little bit more information about what you're using and seeing? For example:
    What restriction enzymes are you using? Did you also include DNA for your control phage? If so, what phage is it? Chidiebere? Could you share a gel image of what you are seeing?

Thanks.
Vic

asprenkle
Anyone have any trouble getting a good RE digest of Gordonia DNA? I've got good DNA from M. foliorum, and we're trying Gordonia rubripertincta for the first time this year. I've noticed it's a bit waxier than M. foliorum. I'm getting OK DNA concentrations with a new Wizard kit, but my Mf DNA cuts where my Gr does not. Any tips/tricks I'm missing?
Thanks!
| posted 19 Nov, 2020 18:06
The only enzyme that cuts well for us in Gordonia rubripertincta is NspI. See attached.
| posted 19 Nov, 2020 18:14
erin.windsor
The only enzyme that cuts well for us in Gordonia rubripertincta is NspI. See attached.

Thanks for sharing this, Erin. We too have found that NspI reliably cuts DNA of phages isolated on G. rubripertincta. We've also found that BamHI, HindIII, and SacII also cut a few of the G. rubripertincta phages we've isolated. Your results will defintiely help us get a better grasp of which enzymes are best to recommend moving forward.

Vic
 
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