By Sabrina Strano
This quantity offers state-of-the-art of tools that may be priceless for either simple and translational researchers to behavior chemoprevention preclinical reports. Written within the hugely profitable Methods in Molecular Biology series layout, chapters contain introductions to their respective subject matters, lists of the mandatory fabrics and reagents, step by step, quite simply reproducible laboratory protocols, and pointers on troubleshooting and warding off identified pitfalls.
Authoritative and functional, Cancer Chemoprevention: tools and Protocols aims to make sure profitable leads to the extra research of this important field.
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Extra resources for Cancer Chemoprevention: Methods and Protocols
1 Schematic representation of the experimental workflow RNA HYBRIDIZATION 851 human microRNAs 18 Sara Donzelli et al. 0. Each slide is an 8 × 15K format (~15,000 features printed in an 8-plex format, eight individual microarrays on a 1″ × 3″ glass slide) printed using Agilent’s 60-mer Inkjet Technology, which, unlike competitive platforms, synthesizes 40–60-mer oligonucleotide probes directly on the array, resulting in high-purity, high-fidelity probes. This miRNA platform requires small input amounts of total RNA—in the 100 nanogram range—because it uses a high-yield labeling method, and does not require size fractionation or amplification steps that may introduce undesired bias during miRNA profiling.
RNA-Seq is also well suited to analyze differential exon usage, gene fusions, and RNA editing. There are several commercial but also free RNA-Seq software packages available. TopHat and Cufflinks for instance are free, open-source software tools that perform the mapping, and transcript assembly and quantification, respectively. The differential expression can be analyzed via Cuffdiff [8, 9]. Take care that the cells are free of mycoplasms. The Ribo- minus protocol removes only ribosomal RNA and retains the whole spectrum of RNA molecules, including RNA from mycoplasms!
Discard the supernatant and wash the beads with 200 μL freshly prepared 80 % ethanol without disturbing the beads attached to the magnet. Wait for 30 s, remove the ethanol and wash again with 200 μL of 80 % ethanol. Air-dry the pellet for 15 min while still being attached to the magnet. 5 μL of Resuspension Buffer. After 2-min incubation 38 Frauke Goeman and Maurizio Fanciulli at RT place the plate on the magnet for 5 min. 3 mL PCR tube. 4. Purify again with AMPure XP Beads by adding 50 μL of beads to the samples.
Cancer Chemoprevention: Methods and Protocols by Sabrina Strano