By Jun Zhang, Gregg Rokosh
This ebook provides either state-of-the-art and proven tools for learning cardiac gene expression. The protocols supply a template for good examine, and canopy the method via screening, research, characterization, and useful affirmation of novel genes or identified genes with a brand new functionality. The concluding component to the publication highlights tools that facilitate overexpression or cardiac-specific precise gene deletion.
Read Online or Download Cardiac Gene Expression: Methods and Protocols (Methods in Molecular Biology Vol 366) PDF
Best biology books
A newly revised version of the traditional reference for the sphere this present day? up-to-date with new phrases, significant discoveries, major scientists, and illustrationsDevelopmental biology is the examine of the mechanisms of improvement, differentiation, and development in animals and vegetation on the molecular, mobile, and genetic degrees.
Environmental version performs a major position in lots of organic and ecological dynamical platforms. This monograph specializes in the research of oscillation and the steadiness of hold up versions taking place in biology. The e-book offers fresh study effects at the qualitative habit of mathematical versions lower than diversified actual and environmental stipulations, masking dynamics together with the distribution and intake of nutrients.
- Overwrites to biology. The speculations of XIX century naturalis
- Molecular Biology and Biotechnology of Plant Organelles: Chloroplasts and Mitochondria
- Advances in Systems Biology
- Biology Education for Social and Sustainable Development
Extra resources for Cardiac Gene Expression: Methods and Protocols (Methods in Molecular Biology Vol 366)
Do not allow the microarrays to dry. 3. Preparation of the Hybridization Cocktail and Hybridization 1. Prepare the hybridization cocktail for a single probe array hybridization as shown in Table 1. The recipe takes into account that it is necessary to make extra hybridization cocktail owing to a small loss in volume during each hybridization. Scale up the volumes for hybridization to multiple arrays. Add the cRNA last. If an error is made, you will not have to redo the fragmentation step. 2. Heat the hybridization cocktail to 99°C for 5 min prior to use.
Preparing the Bioanalyzer The electrodes should be decontaminated before each run. 1. Place a washing chip (electrode cleaner) filled with 350 µL RNaseZap in the Bioanalyzer and close the lid for 1 min. Remove the washing chip from the Bioanalyzer. 2. Place another washing chip filled with 350 µL DEPC-water in the Bioanalyzer, and close the lid for 30 s. 3. Remove the washing chip and leave the lid open for 10 s to allow the electrodes to dry. 3. Loading the Chip Remove the chip from packaging and inspect the under-side wafer of the chip for any defects.
The following steps are carried out at RT. Work quickly through the whole cleanup procedure to minimize the risk of RNA degradation. 22. Add 10 µL β-mercaptoethanol (β-ME) per 1 mL buffer RLT before use. Caution: β-ME is toxic; dispense in a fume hood and wear appropriate protective clothing. Buffer RLT is stable for 1 mo after addition of β-ME. 23. Add 350 µL Buffer RLT to the 100 µL RNA-containing solution, and mix thoroughly. 24. Add 250 µL ethanol (96–100%) to the diluted RNA, and mix thoroughly by pipeting.