By Gary R. Skuse, Maureen C. Ferran
This quantity has been assembled for scientists attracted to uncomplicated and utilized biomedical learn directed towards figuring out the improvement, genetics and serve as of cardiomyocytes. The equipment and protocols contained inside it tackle telephone tradition innovations, cardiomyocyte differentiation and redifferentiation, experimental induction of cardiomyopathies, introducing genes into cardiomyocytes, genomic methods to the certainty cardiomyocytes, cryopreservation of neonatal cardiomyocytes and modeling of cardiomyocyte functionality. Written within the hugely profitable Methods in Molecular Biology sequence structure, chapters comprise introductions to their respective themes, lists of the mandatory fabrics and reagents, step by step, with no trouble reproducible laboratory protocols and pointers on troubleshooting and fending off recognized pitfalls.
Practical and present, Cardiomyocytes: equipment and Protocols explores complimentary components of cardiomyocyte technological know-how that, taken jointly, can tell people with a large diversity of interests.
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Extra info for Cardiomyocytes: Methods and Protocols
Alternatively, an Agilent BioAnalyzer trace could be gained on the amplified material. It is advisable to electrophorese the gel sufficiently to allow good visualization of any material in the 100–120 bp range. Typically, detection of sharp bands in this range indicates the presence of PCR primer-dimers or PCR amplification of core adapters that were not sufficiently removed after the ligation step. If these low-molecular-weight products represent a significant fraction of the total sequencing library, then it is usually desirable to perform a second size-selection step to remove them.
Chlopclkova Sˇ, Psotova J, Miketova P (2001) Neonatal rat cardiomyocytes—a model for the study of morphological, biochemical and electrophysiological characteristic of the heart. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 145:49–55 7. Nuss HB, Marban E (1994) Electrophysiological properties of neonatal mouse cardiac myocytes in primary culture. J Physiol 479(2): 265–279 Chapter 3 Deep Sequencing of Cardiac MicroRNA-mRNA Interactomes in Clinical and Experimental Cardiomyopathy Scot J. Matkovich and Gerald W.
Purify the DNA from the agarose slice using a Qiagen Gel Extraction Kit (see Note 7). Elute in 30 μL EB. 3, sequences are available in Table 1. Components for PCR: DNA from Step A 10 μL (This will retain 20 μL of the gel-purified material from step A, should there be a future need to add a different index sequence, or if optimization of the number of PCR cycles is required. 5 μM 1 μL PCR index-specific reverse primer, 25 μM 1 μL Nuclease-free H2O 12 μL Total reaction volume 50 μL 1. Amplify using the following PCR protocol: 30 s at 98 C.