[DIYbio] Re: Need some help starting a project (Agrobacteria-mediated transformation of a dicot via floral dip)

Hi! AFAIK there already are pCambia Vectors which already contain EGFP unter a plant promoter. So ready to use for your floral dip. 

It may be best to try floral dip for you, because it yields quick results without needing tissue culture.





On Thursday, November 27, 2014 12:03:13 PM UTC+1, antomicblitz wrote:
I'm currently an undergrad studying molecular biology, and I recently joined a community lab after taking biochemistry lab at my school. In my class, we transformed a colony of E. coli using egfp and I thought it was really cool, and I've kind of been obsessed with trying it with other organisms as well, so I wanted to see if maybe I could do the same thing with a plant. After spending a few days at the library and searching through a couple of scientific journals, I found that I could do a similar procedure with agrobacteria, which can use a disarmed tumor-inducing plasmid to inject a segment of tDNA from another binary plasmid into a plant genome (I'm still not 100% sure how that works or where in the genome it gets incorporated, but I know it works at least). I would like to try and put egfp into a plant and have it expressed at detectable levels. I've had people tell me it's not too difficult to do. If any of you have done it before, or if you know what is involved, could you possibly give me some pointers on how/where I can order the appropriate plasmids, and which strain of agrobacteria I can use? I found a couple of protocols, so I have a general idea of what I'm gonna do (floral dip), but I'm still a bit iffy on how to select the right plasmids and bacteria, and where exactly on the binary plasmid I should ligate my gene of interest. Any help is much appreciated :).

--
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diybio@googlegroups.com. To unsubscribe from this group, send email to diybio+unsubscribe@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+unsubscribe@googlegroups.com.
To post to this group, send email to diybio@googlegroups.com.
Visit this group at http://groups.google.com/group/diybio.
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/9fe97591-8974-413b-b6b2-d487de903331%40googlegroups.com.
For more options, visit https://groups.google.com/d/optout.

  • Digg
  • Del.icio.us
  • StumbleUpon
  • Reddit
  • RSS

0 comments:

Post a Comment