Hi,
I have a problem regarding to identifying a transformed Arabidopsis seeds on MS media. I have used Hygromycin B (conc. 20 ug/ml) as a selection marker. I have been growing seeds and they grow fine (all green). However, within 2 weeks all Arabidopsis turn to be white. Do you guys have an idea how to identify a transformed seed?
Maybe useful for you, Have a look
https://www.scionresearch.com/__data/assets/pdf_file/0004/59503/NZJFS2731997WAGNER280-288.pdf
Did you transform using floral dip method? If so, you could repeat the infection procedure the following week to increase transformation efficiency.
And for screening purpose you could increase the drug concentration to 25 or 30mg/l. You should be able to identify the healthy transformed seedlings within 8-10 days post germination.
Faiza Nizamani and Hirotada Fukushige
, thanks your recommended articles.Papri Basak , Yes I had actually transformed using floral dipping. I had also repeated floral dipping the following week.
Hi,
Floral dip methods using hygromicyn B as selection marker, you may select in the four first days of growing. Use the length of hypocotyls to select hygromicyn-resitant. Please, the article below can be helpful for you
Hi,
The agrobacterium you transformed should be identified the interest one by PCR. There are some negative agrobacteriums existing in the screen plate with resistance.
Hyg resistance seedlings should grow normally even after two week. If you are not getting this I would say your transformation did not work. Selection of transformed seedling on hyg plate can be done at one week old seedling. The resistance seedling have normal root and vegetative growth, however the susceptible seedlings have much reduced root and shoot growth.
- Badges
- Science topic
- Similar topics
- Agriculture
- Horticulture
- Seeds