[Arabidopsis] Re: Arabidopsis PEG transformation

Abhijit Karve via arab-gen%40net.bio.net (by abhikarve At gmail.com)
Mon Nov 13 14:37:16 EST 2006


Hi ,
I have been with Arabidopsis protoplast for almost 3 years now, from the
limited experience I have. I think to get good protoplasts and transfection
there are several important points.
1. growth of the plant meterial and the nutrition.
2. Isolation of protoplasts
2. the PEG incubation time and the amount of DNA
but to comment on any of that  I need to know what specific things you used
in your experiment.
Abhijit Karve

On 11/13/06, arab-gen-request At oat.bio.indiana.edu <
arab-gen-request At oat.bio.indiana.edu> wrote:
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>    1. PEG-mediated Arabidopsis Transformation (Zhang, Yi)
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> Message: 1
> Date: Fri, 10 Nov 2006 15:05:27 -0800
> From: "Zhang, Yi" <zhangyi At wsu.edu>
> Subject: [Arabidopsis] PEG-mediated Arabidopsis Transformation
> To: <arab-gen At magpie.bio.indiana.edu>
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> Dear All:
>
> Recently, I've been trying to transform Arabidopsis protoplasts with PEG
> method, but failed.
> The plant is 3 week old Col in greenhouse, watered every other day.
> The protoplasts look good under microscope, but I got no transformants
> with 35S::GFP at all.
> I followed the protocol by Abel & Theologis from Arabidopsis Protocols
> (Humana Press, 1998).
> My question is if there is any trick in the transformation procedure?
> Actually, I've tried
> several protocols from different labs, but all the experiments failed.
>
> Does the whole transformation process have to be operated under sterile
> condition?
> Is there any factor considered critical or important for a successful
> transformation?
> I really expect to learn from your advice if you have experience with such
> transformation.
>
> Thank you very much in advance for your helpful reply!
>
>
> Yi
>
>
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> End of Arab-gen Digest, Vol 19, Issue 7
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ABHIJIT KARVE
204 BRC BUILDING
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