By Yurong Liang, Xin Lu, David L. Perkins (auth.), Jun Zhang, Gregg Rokosh (eds.)
Cardiac Gene Expression: tools and Protocols provides either state of the art and proven equipment for learning cardiac gene expression. The protocols supply a template for good examine, and canopy the method via screening, research, characterization, and sensible affirmation of novel genes or identified genes with a brand new function.
Section I, Cardiac Gene Expression Profiling: the worldwide point of view, discusses numerous varied methods to analyzing, deciding on, and studying adjustments in transcriptome gene expression. part II, Cardiac Gene rules: Gene-Specific mRNA dimension within the Myocardium, outlines extra delicate and gene-targeted expression tools. part III, Cardiac Gene legislation: Promoter Characterization within the Myocardium, offers protocols for the research of underlying gene law mechanisms by means of concentrating on the interplay of transcription elements with their cognate cis binding components. part IV, In Silico review of Regulatory cis-Elements and Gene legislation, and part V, Cardiac unmarried community Polymorphisms, emphasize new analytical methods for decoding the useful components buried within the three billion nucleotides of the human genome and different version genomes. The concluding part, Gene Overexpression and concentrating on within the Myocardium, highlights equipment that facilitate overexpression or cardiac-specific precise gene deletion.
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Extra resources for Cardiac Gene Expression: Methods and Protocols
4. Discard the flowthrough, and transfer the spin column into a new 2-mL Collection Tube (supplied). 5. Pipet 750 µL of the cDNA Wash Buffer onto the spin column. Centrifuge for 1 min at ≥8000g (≥10,000 rpm). Discard flowthrough. 6. Open the cap of the spin column and centrifuge for 5 min at maximum speed (≤25,000g) to dry the membrane completely. Discard flowthrough and Collection Tube. 7. 5-mL Collection Tube, and pipet 14 µL of cDNA Elution Buffer directly onto the spin column membrane. 8. Incubate for 1 min at room temperature and centrifuge for 1 min at maximum speed (≤25,000g) to elute.
7 mL SafeSeal tubes (RNase-free; Sorenson, cat. no. 11510). 1 to 10 µL sterile, filter-free pipet tips with sharp tips, not beveled. Lab tape or Tough Spots (Diversified Biotech, T-Spots). 7. Washing, Staining, and Scanning of Affymetrix GeneChip Probe Arrays 1. Water, molecular biology grade (BioWhittaker Molecular Applications/Cambrex, cat. no. 51200). 2. Distilled water (Invitrogen, cat. no. 15230-147). 3. 50 mg/mL BSA solution (Invitrogen, cat. no. 15561-020). 4. R-Phycoerythrin Streptavidin (Molecular Probes, cat.
Transfer the spin column into a new 2-mL Collection Tube (supplied). 6. Pipet 500 µL IVT cRNA Wash Buffer onto the spin column. Centrifuge for 15 s at ≥8000g (≥10,000 rpm) to wash. Discard flowthrough. 7. Pipet 500 µL 80% (v/v) ethanol onto the spin column and centrifuge for 15 s at ≥8000g (≥10,000 rpm). Discard flowthrough. 8. Open the cap of the spin column and centrifuge for 5 min at maximum speed (≤25,000g) to dry the membrane completely. Discard flowthrough and Collection Tube. 9. 5-mL Collection Tube (supplied), and pipet 11 µL of RNase-free water directly onto the spin column membrane.
Cardiac Gene Expression: Methods and Protocols by Yurong Liang, Xin Lu, David L. Perkins (auth.), Jun Zhang, Gregg Rokosh (eds.)