Unformatted text preview: At this point you should see a little cloudy bacteria around the leaf edges. Bacteria should not be growing over the top of the tissue; if it is you used too much inoculum. 6. Transfer tissue to selective medium (DBI with 200mg/l cefotaxime and 50mg/L kanamycin) in 20x100 mm plates. Put plates under lights in growth room. (We will do this for you.) 7. Expect to see shoots after about 10 days. Excise shoots when they are large enough to handle. 8. Test shoots for GUS activity: slice a thin section off the bottom of the stem and submerge the section in GUS (x-gluc) stain at 37 ° C for 4-5 hour or overnight. 9. Transfer shoots positive for GUS activity to rooting medium. DBI media (per Liter) MS salts 4.3g Thiamine 4mg Myo-inositol 100mg Sucrose 30g IAA 1mg Kinetin 2mg pH 5.6 Agar 8g 30...
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- Spring '09
- Surface tension, Plant morphology, Agar, Agrobacterium culture, Start Agrobacterium culture, larger leaf pieces