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Vortex mixer (Scientific Industries, model: Vortex-Genie 2, catalog number: SI-0236) Ensure that the samples do not thaw during crushing. Figure 2. Crushed leaf sample before incubation in the bacterial isolation buffer; into a new 1.5 ml Eppendorf tube by pipetting.
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crushing leaf in eppendorf using vortex Extraction of Genomic DNA from Rice Leaf by using a 5-ml Tube Crush 0.3 g of rice leaf, put it into the 5-ml tube, turn it upside down, and then mix it by Dispense 100 l of TE buffer and completely dissolve it by using a vortex. (25) Pour the mixture into a 1.5-ml tube and store it in a freezer.
Place all leaf sections obtained in a 50 mL tube filled with sterile water and wash thoroughly using a vortex mixer. Repeat this washing operation three times. Spread the sample uniformly over Kimtowel wiping cloth and dry in a dryer set at 65ºC for 3 hours. Grind thoroughly using a pestle or milling machine*3 until the sample becomes powdery.
3. Crush the frozen slice with a P1000 tip. 4. Add 200 ul of TAE with EtBr (EtBr is not necessary but allows monitoring the extraction) 5. Vortex 6. Spin at 14krpm, room temperature, in an eppendorf 5415C microfuge for 1minute. 7. Remove the buffer and save. 8. Repeat 4-7. 9. You may then heat the agarose and the 400ul pooled TAE to 70 degrees C. 10.
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So, probably I was making a mistake in crushing. I think, I was crushing the leaves for a long time in order to make it fine and that was causing problems. Also, I was using between 80-100 mg leaves.
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crushing leaf in eppendorf using vortex Summary. Citrus Greening is a particularly destructive disease affecting citrus crops globally. Presented here is a simple method using PCR and genomic DNA extraction of citrus leaf tissue for the accurate and precise identification of the citrus greening pathogen, Candidatus liberibacter spp. Cite this Article
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6. Vortex and incubate at RT for 10 min. 7. Centrifuge the samples at 13000 rpm for 10min at 4°C to remove extracellular material and glass beads. 8. Transfer the supernatant to new tubes. 9. Add 0.2ml of chloroform per 1ml TRIZOL 10. Shake vigorously by hand for 15 seconds. 11. Incubate at
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Leaf sample was finely crushed using mortar and pestle and allowed to dry at room temperature for about 5 min. Crushed leaf sample with sand (100 mg) was transferred into a 1.5 mL eppendorf tube.