Transforming Arabidopsis is one of the easiest things to do. Easy transformation has certainly helped make Arabidopsis the premier model plant for scientific study. A printable protocol for Arabidopsis Transformation by Agrobacterium floral dip can be downloaded here. A cheap and efficient way to get your binary vector of choice into Agrobacterium is by the Agrobacterium freeze and thaw transformation method.
Notes:
- Other pages on on this website show how to select for transformants on sterile plates.
- Healthy plants are really important for the transformation process to work.
Which genes can we transform using this method
Any gene or piece of DNA can be put into plants provided it fits between the TDNA left and right border.
Dear Rahul,
Could you please upload your protocols for crossing arabidopsis to create double/triple mutants and your RNA extraction + cDNA synthesis for gene expression analysis? That would be very helpful. Thank you in advance.
M. Osman
M. Osman,
At the moment I do not have video protocol for any of those techniques. When I have time I will try to make those videos.
Best,
Rahul
Hi,
I have a question about agro resistances to antibiotics and strains. My colleague has a problem growing spectinomycin vectors in GV3101. What concentration of antibiotics (combination) should be in the LB media? Do you use only antibiotics resistance to get agro colonies or you check presence of your vector by other methods?
Yes, I use antibiotics to select for the binary vector. I have not used spectinomycin binary vectors in agro. 50 ug/ml would be typical concentration. You could use less or more depending on how the agro grows or does not grow. For example, I have used a kanamycin vector in he past that worked better with have the dose of kanamycin (25 ug/ml instead of 50 ug/ml).