By Kan Wang
A most beneficial choice of Agrobacterium-mediated transformation options for state of the art plant genetic engineering, useful genomic research, and crop development. quantity 1 info the main up-to-date thoughts to be had for twenty-six plant species drawn from cereal plants, business vegetation, legume vegetation, and vegetable crops, and offers numerous equipment for introducing DNA into 3 significant version plant species, Arabidopsis thaliana, Medicago truncatula, and Nicotiana. The authors additionally define the fundamental tools in Agrobacterium manipulation and techniques for vector development. quantity 2 includes one other thirty-three confirmed strategies for root crops, turf grasses, woody species, tropic crops, nuts and end result, decorative crops, and medicinal crops. extra chapters offer equipment for introducing DNA into non-plant species, akin to micro organism, fungi, algae, and mammalian cells. The protocols persist with the profitable equipment in Molecular Biology™ sequence structure, each one providing step by step laboratory directions, an advent outlining the foundations at the back of the process, lists of the mandatory gear and reagents, and pointers on troubleshooting and warding off identified pitfalls.
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Extra resources for Agrobacterium Protocols, Second Edition: Volume I (Methods in Molecular Biology Vol 343)
13. Storage of stab cultures (like other methods) depends, in part, on reducing the cell’s metabolic rate. Agrobacterium will survive at room temperature for many months on stabs made with nutrient agar. Yeast-manitol agar is another good choice for stab culture media. Longest survival times occur when storage of inoculated stabs is at 2 to 4°C (3,4). 14. Clear labels and good records are important components of all storage methods. Labels should note the strain name and any plasmids it contains.
5) of AB*I medium increases expression of the vir genes (10). 0 by buffering with 25 mM Bis-Tris rather than MES when a nonacidic defined medium is desired. 7. The selective nature of enrichment media depends primarily on the presence of substances that are toxic to fungi and some bacteria. Agrobacterium forms shiny, white, raised colonies that may become mucoid or turn orange-brown after extended incubation on media 1A and 1E (3). The addition of tellurite at 80 µg/mL for 1A medium and at 160 µg/mL for 1E medium improves selectivity for Agrobacterium, although Rhodobacter and other Rhizobium species are also tellurite resistant (11).
Well-designed sets of plasmids that consists of a binary vector with these sites and high-copy cloning vectors with expression cassettes and subsets of the sites are called as modular vectors (38). 2. The sites for the GATEWAY® system. Molecular cloning based on restriction enzymes and DNA ligases is not always straightforward. The GATEWAY system (Invitrogen) is a cloning technology based on the site-specific recombination system of phage lambda. A step of molecular cloning may be performed in a single tube within a few hours, and E.
Agrobacterium Protocols, Second Edition: Volume I (Methods in Molecular Biology Vol 343) by Kan Wang