Main Menu

Ph.D Thesis

Web links

Job banks

Login Form

Floral dip and fluoresent maker, very simple transformation

Written by nguyen

Friday, 01 May 2009 17:16

Floral dip of Agrobacterium mediated transformation combine with fluorescent marker gene, a very simple method which any lab can do for plant transformation and it is easy than ever. Below is step by step:

1. Growth plant for transformation, 4 plants per pot and two pots are enough to get 50-100 transgenic seeds

2. As plant have flower cut main first flower make plant have more flower and waiting until ready for transformation

3. Depend your pot size, use the plastic or paper cup to hold Argrobacterium solution for dip flower in

4. Dip all flower in the Argrobacterium solution in few seconds

A kid at 8 years old can do that too

5. Lay down pots in the tissue paper to absorb transformation solution and put a glass cover tray to keep humidity

6. After overnight lay down put the pots stand up in the tray

7. Repeat step 4-6 after 4-5 days of previous transformation, do two or three transformations

8. Water plant until the seed almost matured and let them dry

9. Use tray with net that small enough for seed go through

10. Depending the fluorescent of gene use filter and light correspond with it, for DsRed gene use red filter and green light to collect the fluorescent seed

11. Because the Arabidopsis is too small, can use microscope and attached filter and flash light as the picture to get the transgenic seed easily.

12. Through microscope you can see the red seed as below

13. Three different genes yellow, green and red shown as picture below

14. At T2 plants you can see a segregation of red seed and WT in the silique with ratio 3 to 1 if one copy of gene inserted or multi copy inserted but at same loci