AgroBacteria DIY BIO DALLAS

102 views
Skip to first unread message

Josh Melnick

unread,
Oct 31, 2016, 5:16:16 AM10/31/16
to DIYbio
I am trying to start a DIYBIO group here in Dallas at Dallas Maker Space

I am working on Plant Tissue Culture at the moment, I started looking into ordering AgroBacteria, but it seems I need a special licence to buy it across state lines. 
From what I have read the license is quite hard to get? 

Cory Tobin

unread,
Oct 31, 2016, 5:35:13 AM10/31/16
to diybio
For domestic transportation of Agrobacteria you need permission from USDA APHIS which you can request using form PPQ-526.  Plan a month or so in advance because they're kind of slow.

If you plan on transporting plants that were genetically transformed with Agro you'll need USDA APHIS form 2000.  Although, Arabidopsis thaliana is exempt.

-cory

Josh Melnick

unread,
Oct 31, 2016, 9:52:09 AM10/31/16
to DIYbio
Is it hard to get approved? Do I need the form each time? What strain should I get?

Cory Tobin

unread,
Oct 31, 2016, 2:45:13 PM10/31/16
to diybio
Yeah, its a pain. Before you can begin the application process you need a Level 2 eAuthentication account.
https://www.eauth.usda.gov/MainPages/eauthWhatIsAccount.aspx
You may have to show up to a local USDA office in person with ID.

GV3101 is the strain that many people use.

Sebastian S Cocioba

unread,
Oct 31, 2016, 2:54:40 PM10/31/16
to diy...@googlegroups.com
You need an APHIS shipping permit from the USDA which requires a business or institution. Start filing for 501(c)3 non-profit status and use that for your space. 

Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC

--
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diy...@googlegroups.com. To unsubscribe from this group, send email to diybio+un...@googlegroups.com. For more options, visit this group at https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To unsubscribe from this group and stop receiving emails from it, send an email to diybio+un...@googlegroups.com.
To post to this group, send email to diy...@googlegroups.com.
Visit this group at https://groups.google.com/group/diybio.
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/8668cf93-3072-4ec2-8a08-ae903e6eef9b%40googlegroups.com.
For more options, visit https://groups.google.com/d/optout.

Sebastian S Cocioba

unread,
Oct 31, 2016, 2:56:06 PM10/31/16
to diy...@googlegroups.com
My personal favorite is GV3101 from Stan Gelvin's Lab at Purdue. EHA and LA are both decent but I like the aggression of the GV line. 


Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC

--
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diy...@googlegroups.com. To unsubscribe from this group, send email to diybio+un...@googlegroups.com. For more options, visit this group at https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To unsubscribe from this group and stop receiving emails from it, send an email to diybio+un...@googlegroups.com.
To post to this group, send email to diy...@googlegroups.com.
Visit this group at https://groups.google.com/group/diybio.

Josh Melnick

unread,
Nov 2, 2016, 10:45:55 PM11/2/16
to DIYbio
Thanks I am in the process of getting the Level 2 account APHIS account now based on what you told me, how do I get the strain GV3101 from Stan Gelvin's Lab at Purdue?? just email him?

If you were someone who has never worked with Agrobacteria before what would you recommend as a first experiment to try and test successful creation of transgenic plant? Something with tobacco and GUS reporter gene? 

Josh Melnick

unread,
Nov 2, 2016, 10:46:33 PM11/2/16
to DIYbio
Thank you.  I am in the process of getting the Level 2 account APHIS account now based on what you told me, how do I get the strain GV3101 from Stan Gelvin's Lab at Purdue?? just email him?



If you were someone who has never worked with Agrobacteria before what would you recommend as a first experiment to try and test successful creation of transgenic plant? Something with tobacco and GUS reporter gene? 


Johannes Debler

unread,
Nov 6, 2016, 6:05:18 AM11/6/16
to diy...@googlegroups.com

Good day,

when you say GV3101, are you talking about the cured strain without its helper plasmid? Or do you mean GV3101:pMP90 or pMP90RK? It is my understanding that without its helper plasmid the strain is not virulent and can therefore not transfer any DNA to the host plant.

Cheers,

Johannes

Sebastian S Cocioba

unread,
Nov 6, 2016, 8:08:40 AM11/6/16
to diy...@googlegroups.com
GV3101::pMP90RK is what I meant. Thanks for bringing that up! Its genomically resistant to rifampicin and the Ti plasmid has gentamicin resistance. Its a solid strain!


Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC

Johannes Debler

unread,
Nov 7, 2016, 5:48:15 AM11/7/16
to diy...@googlegroups.com

No worries.

I ask because I just read a paper where the authors say they compared 3 different strains of Agro, amongst them GV3101 and mentioned that efficiency with that strain was low. But then in their supplemental data they mentioned they used the cured strain. So after contacting the author and asking him about why they used the cured strain, which would be avirulent and SHOULD have low efficiency, if any at all, they said they said it's because many labs use it, and they were not sure which strain exactly it was (with or without Ti plasmid) and that that information may have been lost over the course of the experiments....

One more question,
http://dna-cloning.com/?q=strais
mentions that pMP90 confers gent resistance and pMP90RK confers kan + gent resistance. From what I read so far though the most common kanamycin resistance genes nptII and nptIII confer resistance to various aminoglycoside antibiotics, so I am not sure what the point of the pMP90RK Ti plasmid compared to pMP90 is. But I would guess that pMP90 carries a different resistance gene which must be specific for gent.

Cheers

Josh Melnick

unread,
Nov 7, 2016, 12:25:25 PM11/7/16
to DIYbio
I have been working on the APHIS level 2 account, but you need to pay $150 for the VS16-3 form to get the bacteria, but if you request it from within the same state, I don't need permission right? I have never done this before, but could I just email a University and ask them to send me some bacteria if they are within the same state? 
Reply all
Reply to author
Forward
0 new messages