> From: kklinger at ceres-inc.com
> Hey everyone...
> I was hoping anyone could help me out here. I'm trying to find an
> uncomplicated protocol to isolate Arabadopsis DNA that is clean enough to
> PCR. The best example that I have been able to find is the Qiagen DNeasy
> kits, but these are way too expensive ($1100 per 6x96plates, ouch!!). I
> searched around on the web and found the Dynabeads DNA-Direct, which in
> theory seems to be perfect for my application: quick, easy, low prep
> no centrifugations, no tube changes. Has anyone used these on
Also, does anyone have any other advice on either protocols or kits?
Thank you guys so much in advance.
>> KC Klinger
I don't know what you are doing with the DNA afterwards, and I am not sure
how many samples you want to process in a day, but quick, easy and cheap
may not give you "workable" DNA.
>From my experiences, I have found that different species / tissue types
require different protocols to get good quality, high molecular weight,
genomic DNA. So you might benefit from trying several different kits /
protocols before you get started to see if you get good amplifiable DNA.
Or get a protocol directly form a lab already working with Arabidopsis.
In my lab I use several different protocols depending on the type of plant
- but I do not work with arabidopsis.
a) QIAGEN DNeasy Plant mini columns
b) a basic CTAB extraction (modified from Saghai-Maroof et al 1984)
c) a nuclear isolation and then DNA isolation for plants high in
polyphenolics or acids (modified from Wang, Wang & Zou 1996)
d) a phenol extraction for tissue high in lipid content (Prince et al 1997
- HORT SCIENCE 32(5):937-939)
I can't remember who is making them, but I saw an advertisement for some
cards where you drop some crude extract onto the card and then elute your
DNA off of the card. Sounds easy - maybe someone else can recall the
Hope this helps
Molecular Markers Lab Technician
USDA Agricultural Research Service
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