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vsoza

USA
18 Posts
Posted - March 02 2020 :  2:05:56 PM  Show Profile  Reply to Topic
Rhododendron DNA extractions
Has anyone had trouble getting good DNA yield from fresh-collected Rhododendron samples? If so, have you found a good protocol that has worked for problematic samples?
Edited by - vsoza on March 02 2020 2:10:43 PM

vsoza

USA
18 Posts
Posted - March 02 2020 :  2:11:46 PM  Show Profile  Reply with Quote  Reply without Quote
We have used the Qiagen DNeasy kit but have used ethanol precipitations instead of column cleanup because DNA in vireyas was not binding to the columns.

quote:
Originally posted by vsoza

Has anyone had trouble getting good DNA yield from fresh-collected Rhododendron samples? If so, have you found a good protocol that has worked for problematic samples?
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JulianaMedeiros

USA
12 Posts
Posted - March 10 2020 :  1:06:05 PM  Show Profile  Reply with Quote  Reply without Quote
We've had good luck with phenol cholorform extractions, although we've only tried it on a few species.
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emilygillespie

USA
1 Posts
Posted - April 29 2020 :  11:24:12 AM  Show Profile  Reply with Quote  Reply without Quote
I've used more Qiagen DNEasy than anything over the past decade with good results. I modified for herbarium materia or silica-dried material according to Drabkova et al (2002?)...though their work was done on Juncus or something.

quote:
Originally posted by vsoza

Has anyone had trouble getting good DNA yield from fresh-collected Rhododendron samples? If so, have you found a good protocol that has worked for problematic samples?
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JulianaMedeiros

USA
12 Posts
Posted - April 30 2020 :  09:59:07 AM  Show Profile  Reply with Quote  Reply without Quote
Glad you both like the Quiagen DNEasy, because I have a bunch of those purchased that Im planning to use on R. minus
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RhodoRyan

USA
2 Posts
Posted - May 14 2020 :  5:14:25 PM  Show Profile  Reply with Quote  Reply without Quote
I haven't had a lot of issues with extraction, but I know a member of our lab had some issues with Qiagen.

We most recently used this kit for Target-Capture sequencing with great results:

https://www.asone-int.com/product/invisorb-spin-plant-mini-kit-2/


If you want a cheaper route, we have some CTAB protocols that also worked well for troublesome samples. Email me and I can send them over.

-Ryan
Edited by - RhodoRyan on May 14 2020 5:18:52 PM
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