By Julio Salinas, Jose J. Sanchez-Serrano
This number of comfortably reproducible Arabidopsis protocols has been up-to-date to mirror contemporary advances in plant biology, the of completion of the Arabidopsis genome series, that's crucial for learning plant functionality, and the advance of entire platforms methods that permit worldwide research of gene expression and protein and metabolite dynamics. The authors have incorporated approximately all strategies built in Arabidopsis, others lately tailored from the conventional paintings in crop species, and the latest ones utilizing Arabidopsis as a version procedure. 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 resources for Arabidopsis Protocols, 2nd Edition (Methods in Molecular Biology)
The classical approaches of plant DNA isolation are often designed to produce large amounts of DNA of high molecular weight with sufficient purity. Since the polymerase chain reaction (PCR) 1s the method of choice in modern plant genetic analyses such as map-based breeding and positional gene clonmg, many recently developed protocols are aimed at rapid extraction of a minute quantity of plant DNA that 1ssultable for PCR. This allows large numbers of samples to be analyzed m a short period of time.
Add 600 pL of lsopropanol, vortex briefly, and mnnedlately centrifuge at 18,OOOg for 5 mm m the mtcrocentnfuge. g Carefully remove the supernatant using a Pasteur plpet, leaving the pellet m the bottom of the tube h. Dry the DNA pellet m a Speed-Vat to remove any of the remaining liquid and resuspend m 100 pL of sterile deionized water. 58 Li and Chory 2. Since most plant DNA preparations are often contaminated by polysaccharides, dark adaptmg the plants for 2-3 d before collecting tissues is usually enough to reduce their content m plants.
These methods are equally effective for the Arabzdopszs ecotypes Columbia, C24, RLD, and WaSSllJeWSkiJa. 2. 1. Materials and Equipment 1. Dry seeds of Arabldopsls 2. 1% Trlton X-100 3. Sterrlrzed double-distilled water. Edited From Methods III Molecular Bology, Vol 82 Arabrdopsrs by J Martmez-Zapater and J Salmas 0 Humana Press 35 Protocols Inc , Totowa, NJ Mathur and Koncz 36 4. 5. 6 7 8. 9. 10. 11 12 13. 14. 15. 16 250 mL Sterile Erlenmeyer flasks 0 22 pm Filters for sterilization of solutions.
Arabidopsis Protocols, 2nd Edition (Methods in Molecular Biology) by Julio Salinas, Jose J. Sanchez-Serrano