Please I need to know how to add hygromycin B to my hydroponic solution for transformant selection. My vector construct contain Hygromycin ressistant gene (HPT).
I know how to use the MS selection medium. But right now I'm employing the use of hydroponic solution to grow my transformed peanut plants. I used this method because it is impossible to select transformed plants using soil media. The use of germination bag makes it easier to see the formation of the hairy root and also help for easy phenotype of the root.
I know how to make stock solution of hygromycin B, I need to know how to apply it to the hoagland solution in the germination bag. such that the volume of the Hygromycin won't be too much.
As you all know that the antibiotic is very expensive.
Hint: I have about 150 plants in the growth chamber for this batch of experiment now.
Thank you
Dear James Seun Maku
Pls. find the attached files
http://onlinelibrary.wiley.com/doi/10.1111/j.1467-7652.2005.00138.x/full
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3610267/
http://www.nature.com/articles/srep24941?WT.feed_name=subjects_plant-biotechnology
https://plantmethods.biomedcentral.com/articles/10.1186/1746-4811-9-4
http://www.plantphysiol.org/content/143/1/425.full
Page 415 in: Leslie Wilson, Paul T. Matsudaira, David W. Galbraith, Don P. Bourque, Hans J. Bohnert .1995. Methods in Plant Cell Biology, Academic Press,Pp.555 ISBN. 0080859488, 9780080859484
Hoping this will be helpful
Regards
Prof. Houda Kawas
Thank you Prof. Houda Kawas. Your paper is helpful, I am thinking of a way to utilize the 0.5ml enpendorf tube removing its bottom and then filled the bottom with agar (selection media) so that the hairy root can grow through the agar if it's a true positve transformant. It looks like its going to be a difficult task because I have my plants in a germination bag. But I will sure think on what to do.. If you also have any suggestion you can also share. thanks
You most welcome
that science try and modifies, then will be your methods
I'll send any new idea when I get it sure
Best of luck,
Houda
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