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

Anne Britt via arab-gen%40net.bio.net (by abbritt from ucdavis.edu)
Mon Aug 29 15:04:41 EST 2016


looking at root growth is very helpful


> On Aug 29, 2016, at 10:23 AM, Raghu Badmi <hiraghuraman from gmail.com> wrote:
> 
> 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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>> Today's Topics:
>> 
>>   1. Selection of transgenic Arabidopsis line in       Hygromycin medium
>>      (MAHMUDUL HASSAN)
>> 
>> 
>> ----------------------------------------------------------------------
>> 
>> 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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