Read e-book online Arabidopsis Protocols, 2nd Edition (Methods in Molecular PDF

By Julio Salinas, Jose J. Sanchez-Serrano

ISBN-10: 1588293955

ISBN-13: 9781588293954

This selection of effortlessly reproducible Arabidopsis protocols has been up-to-date to mirror contemporary advances in plant biology, the of entirety of the Arabidopsis genome series, that's crucial for learning plant functionality, and the improvement of entire platforms techniques that permit international research of gene expression and protein and metabolite dynamics. The authors have integrated approximately all suggestions built in Arabidopsis, others lately tailored from the conventional paintings in crop species, and the latest ones utilizing Arabidopsis as a version procedure. Highlights contain the latest methods-transcriptomics, proteomics, and metabolomics - and their novel purposes (phosphoproteomics, DNA microarray-based genotyping, excessive throughput metabolite profiling, and single-cell RNA).

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Extra resources for Arabidopsis Protocols, 2nd Edition (Methods in Molecular Biology) (Не полн.)

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Highly purified mitochondrra are important as a source of many biochemrcal, physiologrcal and molecular applications. The preparation of mitochondrta as a prerequrstte to analyze mttochondrral DNA 1s described in Chapter 12. Purified mrtochondria are also required for protein and RNA import studies (1,2), enzymatic activity measurements (3,4), in organello protein analysis (5,6), to prepare protein extracts and protem complexes from mrtochondrral membranes (7) and for the tsolatron of mitochondrial RNA (8).

The phenol/chloroform extraction is also often used to remove contaminated polysaccharides from the DNA preparation These carbohydrates, which coprecipitate with DNA during alcohol precipitation, inhibit many enzymes commonly used m molecular clonmg procedures and affect electrophoresis during further uses of DNA. Many other methods have been developed to remove these contaminated compounds from DNA preparations including CTAB extractton (3,4), high-salt treatment (5), DEAE-cellulose method (6), Sephacryl S- 1000 chromatography (7), and Caylase M3 digestion (8) 12 Addition of 3M sodium acetate before the phenol/chloroform extraction, rather than at step 11, can result m a better recovery of DNA in the aqueous phase during the organic extraction (9).

2. 4 (with 5M KOH), autoclave, and store at 4°C. Before use, add dtthiotreitol (DTT) to a final concentration of 10 mA4, bovine serum albumin (BSA) to 2 g/L and polyvmylpyrrolidon (PVP) to 6 g/L 7. 2 (with 5M KOH), autoclave, and store at 4°C. Before use, add BSA to 1 g/L. 8. Resuspension buffer ( 1X R) ( 1 L): 400 mM manmtol, 10 mM trtcme-KOH pH 7 2, EGTA-KOH pH 7 2. AdJust pH to 7 2 (with 5M KOH), autoclave, and store at 4°C 9. 5Msucrose, 50 mMMOPS AdJust pH to 7 2 with 5M KOH solution, autoclave, and store at 4°C 10.

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Arabidopsis Protocols, 2nd Edition (Methods in Molecular Biology) (Не полн.) by Julio Salinas, Jose J. Sanchez-Serrano

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