Fw: Quick, easy & cheap plant DNA isolation???

Deanne Bell dbell at qnis.net
Tue Jan 30 12:15:21 EST 2001


> 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
time,
> no centrifugations, no tube changes.  Has anyone used these on
Arabadopsis?
 Also, does anyone have any other advice on either protocols or kits?
 Thank you guys so much in advance.
> 
> KC Klinger

Hi KC
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
manufacturer.

Hope this helps
Deanne Bell
Molecular Markers Lab Technician
USDA Agricultural Research Service
2021 S. Peach Ave.
Fresno, CA 93727
e-mail: dbell at qnis.net
phone: 559 453-3170
fax: 559 453-3088




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