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[Arabidopsis] Re: Arab-gen Digest, Vol 136, Issue 7

Raghu Badmi via arab-gen%40net.bio.net (by hiraghuraman from gmail.com)
Mon Aug 29 12:23:52 EST 2016


Dear Mahmud Hassan,

My protocol:
#Hyg-25mg/L
#plate seeds and incubate 4c dark 2-3 days
#transfer plates to light for atleast 8 hours
#transfer back to dark at 22-24C for atleast 4 days
# look at the seedlings- you should see the elongated hypocotyl for
transformants and others will have short hypocotyl and look dwarf.

I hope this would be of some use to you.

Good luck,

Raghu Ram

On Aug 29, 2016 13:04, <arab-gen-request from oat.bio.indiana.edu> wrote:

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>    1. Selection of transgenic Arabidopsis line in       Hygromycin medium
>       (MAHMUDUL HASSAN)
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> ----------------------------------------------------------------------
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> Message: 1
> Date: Sun, 28 Aug 2016 02:08:54 +0000 (UTC)
> From: MAHMUDUL HASSAN <mahmudul_bau from yahoo.com>
> Subject: [Arabidopsis] Selection of transgenic Arabidopsis line in
>         Hygromycin medium
> To: "arab-gen from net.bio.net" <arab-gen from magpie.bio.indiana.edu>
> Message-ID: <1727940525.844985.1472350134556 from mail.yahoo.com>
> Content-Type: text/plain; charset=UTF-8
>
> Hi All
> I am a PhD student. I have been trying to select transgenic Arabidopsis
> line in hygromycin medium for the last couple months. I followed this paper
> (A rapid and robust method of identifying transformed Arabidopsis thaliana
> seedlings following floral dip transformation, Plant Methods. 2006. 2:19.
> DOI: 10.1186/1746-4811-2-19) but failed to select transgenic plants. Both
> untransformed (Col-0) and Hyg positive plants look like same in the
> selection plate. I used various concentration of Hyg (15, 20, 25) but all
> the time I was unable to differentiate between the susceptible and
> resistant plants. I am using Hyg from Sigma and it was bought recently. I
> am desperately looking for a working protocol for transgenic Arabidopsis
> selection in Hygromycin media.
>
> I would be really grateful if anyone can help me in this regard by giving
> some suggestion for transgenic Arabidopsis selection in Hyg media and
> hopefully any protocol if possible?
> Thank you all
> Regards
> Mahmud Hassan
>
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> End of Arab-gen Digest, Vol 136, Issue 7
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