By Jose M Martinez-Zapater, Julio Salinas
This entire number of present and crucial protocols includes many simply reproducible equipment built to be used with Arabidopsis - a approach for coming near near primary questions in plant biology. The equipment diversity from the fundamentals of starting to be those crops to classy gene cloning techniques and will, in lots of circumstances, even be utilized to different plant species with minor transformations. Sections on genetics, transformation and gene expression research which are particularly useful to scientists occupied with mutant research or generating and examining transgenic crops.
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Extra resources for Arabidopsis Protocols
Quassollo G, Wojnacki J, Salas DA, Gastaldi L, Marzolo MP, Conde C, Bisbal M, Couve A, 9. 10. 11. 12. 13. Cáceres A (2015) A RhoA signaling pathway regulates dendritic Golgi outpost formation. Curr Biol 25:971–982 Kaech S, Huang CF, Banker G (2012) General considerations for live imaging of developing hippocampal neurons in culture. Cold Spring Harb Protoc 201:312–318 Maccioni HFC, Quiroga R, Spessott W (2011) Organization of the synthesis of glycolipid oligosaccharides in the Golgi complex. FEBS Lett 585:1691–1698 Giraudo CG, Rosales Fritz VM, Maccioni HF (1999) Ga2/Gm2/Gd2 synthase localizes to the trans-Golgi network of cho-k1 cells.
Combine Lipofectamine and DNA mixtures and incubate for 20 min at room temperature. While incubating, gently recover original culture medium, wash each well twice with 500 μL Neurobasal Medium per well, and add 1 mL of Neurobasal Medium for transfection (see Note 5). 5. After incubation, add 200 μL of transfection mix to each well and return to a CO2 incubator at 37 °C for 1 h. 6. 22-μm membrane, and return both filtered medium and Neurobasal Medium to 37 °C. 7. After 1 h of transfection, gently wash each well twice with Neurobasal Medium and add 1 mL of filtered original medium plus fresh neuronal maintenance medium.
Turn on the spinning- disk confocal microscope, set the temperature to 37 °C using a temperature controller connected to a heating device, and wait until the temperature is stabilized to 37 °C. Pre-warm the oil used for objectives to 37 °C. 2. In a vertical laminar flow hood, remove the coverslip from the plate using sterile fine forceps, place the coverslip in a 35-mm dish type magnetic chamber for incubating 18-mm round coverslips during live-cell imaging, and add 500 μL of the original medium.
Arabidopsis Protocols by Jose M Martinez-Zapater, Julio Salinas