By Jérôme P. Fennell, Andrew H. Baker
A suite of recent and crucial molecular recommendations for cardiovascular study. those conveniently reproducible equipment variety extensively from generating congenic, consomic, transgenic, and knockout versions of high blood pressure to the gene move of particular genetic fabric utilizing nonviral (polymers, liposomes, and antisense brokers) and adenoviral vectors. extra options defined comprise unmarried nucleotide polymorphism (SNP) genotyping, RNA interference, microarray research, pharmacogenetics, and pharmacogenomics for the genetic dissection of high blood pressure, in addition to a pragmatic technique for deriving cardiomyocytes from embryonic stem cells that might function alternative cells for these broken by means of high blood pressure or center assault. The booklet deals either beginner and skilled high blood pressure researchers an essential number of without difficulty reproducible innovations for profitable examine, paintings that has already dramatically better the outlook for hypertensive sufferers, and provides a lot destiny luck.
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Additional resources for Hypertension: Methods and Protocols (Methods in Molecular Medicine)
46 Popova et al. 11. Remove the pipet from the oviduct. 12. Gently place the fat pad back into the body cavity, stitch the wound, and repeat this procedure on the other side (see Note 19). 13. Operated animals are maintained under optimal husbandry conditions. 6. Identification of Transgenic Animals Integration of the transgene into the recipient genome can be determined by transgene-specific polymerase chain reaction (PCR) or Southern blot with genomic DNA isolated from ear or tail biopsies of the offspring after weaning (Fig.
5. Sometimes several primer pairs have to be tested for the diagnostic PCR. 6. If medium becomes yellow, change it up to twice daily. 7. Before microinjection of positive clones, verify the correct genotype by Southern blot. 8. Alternatively, quite suitable pipets can be purchased from commercial suppliers 9. When the pipet is clogged, flush it with mineral oil and recoat with PVP solution. Acknowledgments The author would like to thank Vanessa Merino, Natasha Alenina, Cibele Campos Cardoso, Cécile Cayla, Tanja Schmidt, and Claudia Wilhelm for helpful discussions.
25. , Maandag, E. , and Berns, A. (1992) Highly efficient gene targeting in embryonic stem cells through homologous recombination with isogenic DNA constructs. Proc. Natl. Acad. Sci. USA 89, 5128–5132. 26. 26 van Deursen, J. and Wieringa, B. (1992) Targeting of the creatine kinase M gene in embryonic stem cells using isogenic and nonisogenic vectors. Nucl. Acids Res. 20, 3815–3820. 27. , Chai, G. , Baltas, L. , et al. (2000) Smooth-muscle contraction without smooth-muscle myosin. Nat. Cell Biol.