By Julio Salinas, Jose J. Sanchez-Serrano
This choice of simply reproducible Arabidopsis protocols has been up to date to mirror fresh advances in plant biology, the finishing touch of the Arabidopsis genome series, that's crucial for learning plant functionality, and the improvement of complete platforms techniques that let worldwide research of gene expression and protein and metabolite dynamics. The authors have integrated approximately all ideas built in Arabidopsis, others lately tailored from the normal paintings in crop species, and the latest ones utilizing Arabidopsis as a version approach. Highlights comprise the latest methods-transcriptomics, proteomics, and metabolomics - and their novel functions (phosphoproteomics, DNA microarray-based genotyping, excessive throughput metabolite profiling, and single-cell RNA).
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Additional info for Arabidopsis Protocols, 2nd Edition (Methods in Molecular Biology)
Pair of forceps. 25, 50-, and 100~pm Sieves. 12 mL Screw-capped glass centrifuge tubes. 10 and 25 mL Sterile pipetes. Pipet pump Pasteur pipets with cotton plugs and teats Hemocytometer Inverted microscope 55 and 90 mm Petri dishes. All glassware should be sterilized prior to use and all operations are carried out usmg a sterile bench. 2. 8 with 1M KOH or 1M HCl. Protoplast medium (PM), and the enzyme solutron are filter sterilized. Growth regulator stocks (1 mg/mL) are filter sterilized and added separately to sterilized media.
The soft transition from one osmoticum to another results m less protoplast bursting and loss between the steps. 20 If the number of protoplasts exceeds lOO/square of the hemocytometer, dilute the protoplast suspension 2 1. Protoplasts embedded m algmate may be slower m dividing by 1 or 2 d. 22 Depending on the quality of protoplast preparation, up to 75% of protoplasts survtve and 2040% of cells will undergo divisions during the first 3-5 d of culture In liquid medmm, an mmal high density may result in aggregation and collapse of protoplasts.
5 Ltlley, R. MC C , Fitzgerald, M. , Rienits, K. G. (1975) Criteria of intactness and the photosynthettc activity of spinach chloroplast preparations. New Phytol 75, l-l 0 6 Deheu, T , and Walker, D A ( 1972) An improved cathode for the measurement of the photosynthettc oxygen evolutton by isolated chloroplasts New Phytol. 71,201-225 8 Purification of Mitochondria from Arabidopsis Mathieu Klein, Stefan Binder, and Axe1 Brennicke 1. Introduction Plant cells contam different subcellular compartments, which serve distinct phystologrcal functions.