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Recent Activity
Gordonia DNA not cutting
| Link to this post | posted 19 Nov, 2020 16:12 | |
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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! |
| Link to this post | posted 19 Nov, 2020 16:58 | |
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Hi Amy, Would you mind sharing a little bit more information about what you're using and seeing? For example:
Thanks. Vic asprenkle |
| Link to this post | posted 19 Nov, 2020 18:06 | |
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The only enzyme that cuts well for us in Gordonia rubripertincta is NspI. See attached. |
245Kb| Link to this post | posted 19 Nov, 2020 18:14 | |
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erin.windsor 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 |
| Link to this post | posted 30 Nov, 2020 14:25 | |
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I was finally able to get cutting, and with NspI but not HindIII. I did not use other enzymes after Dan suggested he only needed to see UC DNA, so I was being frugal with enzymes and gel lanes. I had omitted the 65C incubation after DNA isolation. That is clearly a necessary step  Thanks for your help! |