Agrobacterium transformation protocol

    • [DOC File]Tri-parental Matings

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      1. Introduction of binary constructs into Agrobacterium. Transform the binary plasmid into Agrobacterium tumefaciens strain EHA101 (Selection plates are LB agar with 100 mg/L spectinomycin, 50 mg/L kanamycin and 25 mg/L chloramphenicol), re-isolate plasmid and check by restriction enzyme digestion. 2. Agrobacterium-mediated transformation of Maize

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    • LBA4404 ElectroCompetent Agrobacterium | ig™ | Intact Genomics

      Preparing Camelina sativa for agrobacterium transformation. Required Materials: 30-35 Day old . Camelina. sativa. plant. Scissors (optional) Using scissors, or your fingers, snip/pluck off any flowered or flowering buds from the . Camelina. Remove any opened, or opening, buds of the plant.

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    • [DOC File]Large-scale fluorescent tagging of full-length genes to ...

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      Figure 2 - Potential of oat tissues for Agrobacterium transformation. Tissues of genotype Melys were transformed with AGL1/pSoup/pAL156 (A - experiment AT54) or AGL1/pAL155/pAL156 (B- experiment AT43) according to the protocols described in Table 1. Figure 3- Potential of oat genotypes for Agrobacterium transformation.

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    • Michigan State University

      "Recipient" = Agrobacterium or Rhizobium, often grown on LB at 28oC, or . Pseudomonas or Xanthomonas, often grown on NYGA at 28oC. Selection will often involve two antibiotics. One marker is carried on broad host-range plasmid, such that all helper and recipient strains that have not received this plasmid can be killed. The other marker is ...

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    • [DOC File]Research and Development

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      Agrobacterium cells carrying a binary vector are grown for 24 h in Luria-Bertaini (LB) media supplement with the appropriate antibiotics on an orbital shaker at 28 ºC and 250 rpm. The cells are pelletted by centrifugation at 4000 rpm (1,992 RCF) for 30 min and then re-suspended in an Agrobacterium MS-IM containing 200 ul AS at an OD600nm of 0 ...

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    • [DOC File]Flow chart p - Harvard University

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      Rice transformation procedures. The constructs (all contain hygromycin-resistant gene) were introduced into Agrobacterium tumefaciens strain EHA105, and then transformed into embryogenic calli induced from rice Nipponbare (Oryza sativa L. ssp. japonica) mature seeds. Rice transformation, tissue culture and plantlets growth procedure were mainly ...

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    • [DOC File]Transformation of Aspen INRA 717-1B4 with alcohol ...

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      Transformation and storage of VIGS agrobacteria: Once your silencing vector sequence has been verified, transform a sample into electrocompetent agrobacteria, strain GV3101 (or try EHA105). Before transformation verify that the electroporator, electrocompetent agrobacteria cells, and appropriate selective plates are available.

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    • [DOC File]Purdue University - Indiana's Land Grant University

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      Seedlings were injected rather than dipping cotyledonary petioles (as in the transformation protocol) and the site of infection was therefore kept clear of the culture medium, so that the tissue culture blackening response observed in the regeneration trials was not mistaken for a hypersensitive response to Agrobacterium infection.

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