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[Arabidopsis] Selection of transgenic Arabidopsis line in Hygromycin medium

John Turner (BIO) via arab-gen%40net.bio.net (by J.G.Turner from uea.ac.uk)
Mon Aug 29 03:39:46 EST 2016


Hi

The phenotype is very weak, and the reason why we turned to kanamycin resistance as the marker for transformations. You should see shorter roots in the wild type (test plus versus minus hygro on wild type). If you dont, then hygro is not working. Is light sensitive.

John


John Turner
Emeritus Professor
School of Biological Sciences
University of East Anglia
Norwich Research Park
Norwich NR4 7TJ
England, UK

Mobile Phone 07411429440
email j.g.turner from uea.ac.uk<mailto:j.g.turner from uea.ac.uk>; johngturner2009 from gmail.com


________________________________
From: arab-gen-bounces from oat.bio.indiana.edu <arab-gen-bounces from oat.bio.indiana.edu> on behalf of MAHMUDUL HASSAN <mahmudul_bau from yahoo.com>
Sent: 28 August 2016 03:08:54
To: arab-gen from net.bio.net
Subject: [Arabidopsis] Selection of transgenic Arabidopsis line in Hygromycin medium

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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