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Lab 03-cauliflower_coleus - Lab 3b Producing Plantlets from...

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Lab 3a Production of Plantlets from Flower Organs of Cauliflower Protocol: 1. Outside of the hood, select a clean floret from a fresh cauliflower head. Using forceps and scalpel, trim off 12-15 small pieces of florets for explants 2. In the flow hood, transfer the material into a sterile beaker containing a stir bar 3. Add 70% EtOH and stir for 5 min. Pour off EtOH 4. Add 20% bleach + Tween 20 and stir for 5 min. Pour off Bleach 5. Rinse tissue 3X with Sterile water (1-2min each) 6. Transfer the explants into 2 petri plates containing MS medium AND 2 petri plates containing MS medium supplemented with 2.5 mg/L of kinetin. 7. Seal the plates, label them and maintain them in the light. 8. Observe plants weekly 9. When shoots develop, in 6-8 weeks, separate from mass at base of the shoots and put them into hormone free medium. Roots should form in 2-3 weeks.
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Unformatted text preview: Lab 3b Producing Plantlets from Coleus leaves 1. Leaves from Coleus plants have been collected and provided to you. It is your objective to regenerate shoots that are of a single color – how would you go about doing that with these Coleus leaves? 2. In the flow hood, transfer the material into a sterile beaker containing a stir bar 3. Add 70% EtOH and stir for 3 min. Pour off EtOH 4. Add 20% bleach + Tween 20 and stir for 5 minutes Maximum. Pour off Bleach. 5. Rinse tissue 3X with Sterile water (1-2min each) 6. Lay a leaf out on the inside of a large petri plate. Cut the leaf into 8-10 pieces. 7. Transfer the explants into 2 petri plates containing MS medium plus 2 mg/l BA & 1 mg/l NAA. 8. Seal the plates, label them and maintain them in the light. 9. Observe plants weekly....
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