Transforming Agrobacterium by the freeze thaw method
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- čas přidán 13. 09. 2024
- This video shows you how to transform Agrobacterium with plasmids by freezing and thawing the cells. The freeze thaw transformation method is performed in a single tube and is cheap and efficient. Additionally, the competent cells are very easy to prepare. A printable protocol can be downloaded here patharkar.com/...
A print protocol and more details to go with the video can be downloaded here: patharkar.com/2018/11/transforming-agrobacterium-by-the-freeze-thaw-method
Hello! can you re-upload the document please
@@ruiznery Done! You probably would have to reload the page a couple of times in your browser to get it. Let us know if you have any more questions. Gracias por contactarnos.
@@catherineespinoza5917 Muchísimas gracias!! Me ayuda bastante en mi proyecto, cuando acabe les adjunto, voy a utilizar su método. Bendiciones y que tengas un excelente 2021
Print protocols and more background can be found on patharkar.com
thank you Rahul
You are welcome. All the best.
Hi, Rahul. Long time no see. Thanks, great video.
Great to hear from you! It has been a long time! All the best to you.
Hi its really helpful but can you pls manage background noice
Unfortunately it is filmed in a lab with a fume hood and other equipment that makes noise. How would you suggest to handle this background noise?
Hi, I am looking to transform Agrobacterium I have, and your procedure says to use "standard miniprep binary plasmid." Is there a way I can synthesize this or buy this? What are my alternatives here without spending lots of money?
Also, I eventually want to transform Arabidopsis via floral dip, but I want to be able to confirm it was successful. Is there a DNA marker that could be revealed chemically or through PCR that could be used to confirm a successful transformation? How can i confirm that the process was a success?
If you have access to a lab, my miniprep procedure will cost you very little (about $0.05/sample) to prepare the DNA. See my miniprep procedure patharkar.com/2019/03/plasmid-dna-isolation-improved-yield-no-kit. In general you will have to prepare your plasmid DNA or pay a service to prepare it for you. Paid services might charge $50-100 to prepare the DNA. DNA synthesis will cost even more. You have options based on our goal.
Hi, just wondering how long can the agrobacterium lasts after snap frozen and stored at -80 degree?
I have used 4 year old frozen cells and did not notice any decline in the number of colonies. Therefore they last greater than four years.
@@RahulPatharkar I see. Thank you.
thank you Rahul
You are welcome. All the best.