DNA extraction, high polyphenol - help (Oct/14/2005 )

hi..

i extracted DNA from plant. i use leaves, but i didnt get DNA on agarose gel (electrophoresis).
but, i get ratio A260/A280 1,7 ; 1,3 ; 2,1. I use CTAB and PVP (40.000). Some body, can help me?
why i didnt get DNA......

thank you

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Dear friend,
What is your amount of you extracted DNA. You might get something very pure but too little to view from gel.

Could you descript you protocol of plant DNA extraction?

Thank you

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oh i see...may be..it's too little cocentration.
i use 5 uL DNA and 2 uL loading dye.
protocol DNA extraction by Doyle&doyle modified by Lodhi (1994)
Fresh leaves grinding with liquid nitrogen add buffer extract (Tris-Cl, B merkaptoetanol, PVP, NaCl)
when i sentrifuge (+chloroform:iaa) there is 3 phase.
and debris phase are so cloudly and viscous.

but, i still confuse
i have 3 time for phenol extraction but it wasn't purity
A 260/A280 still same.
what happen.....?
thank you

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Dear freind,

How much starting material use use to extract your DNA?
By the way it shouldn't be so little.

Bellow is the web of side of university of Zurich, institute of plant biology.
They have some good protocol. Try it out

http://www.unizh.ch/botinst/Cyto_Website/s...A/totalRNA.html



Best regards

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Dear friend,
Look at this webside as well.

http://www.pa.ipw.agrl.ethz.ch/research/Ap...ls/ctab-xtr.htm


Best regards

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thank you very much
i'll try extract again....

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[quote name='tomoe' date='Oct 19 2005, 04:14 AM' post='28319']
[quote post='28211' date='Oct 18 2005, 07:40 AM' name='Hadrian']
Dear friend,
Look at this webside as well.

http://www.pa.ipw.agrl.ethz.ch/research/Ap...ls/ctab-xtr.htm


Best regards
[/quote]
Hi...
today, i try running my result DNA on 0,8% agarose.
I choose DNA which are has high purity( 1,7- 1,83) and
I use 10 microlitre DNA sample with 4 microlitre dye.
Cocentration 60-85 microgram/microlitre.
it means 0,6-0,85 ug on one coloum

but, i still didnt find bind on my agarose gel.
Any sugestion about it?

I still confuse....
it is different result if i use miniscale DNA extraction?
Protocol which i read, use largescale DNA extraction

Thank You....

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