Fspi restriction enzyme

Unless indicated otherwise, experiments were based on isolated AMs or atrial tissue from the hearts of different species: (a) 12–16 weeks old female mice in the C57BL/6N background; (b) 16–20 weeks old female Wistar rats; (c) female New Zealand White rabbits 17 weeks old with a body weight of 3.0 to 3.5 kg; and (d) 3-month-old healthy German landrace pigs of either gender with body weights of 40–45 kg. Mice and rats were euthanized by cervical dislocation under 1.5–2% isoflurane anesthesia; rabbits by pentobarbital (400 mg/kg). The generation of Ryr2^{S2808A/S2808A} knockin mice was described previously (Lehnart et al., 2005 (link)); for genotyping we used the following primer pairs: F 5′-ATCCCGAGGTAATCAGGTTTCT-3′, R 5′-AGTTGGGTTCAAAGTTCTAGGC-3′; and PCR product digestion by Fsp I restriction enzyme (New England Biolabs). This study was carried out according to guidelines for the care and use of laboratory animals following directive 2010/63/EU of the European Parliament and the Council of the European Union, Strasbourg, France, and in keeping with NIH guidelines. After mouse, rat, or rabbit euthanasia, the heart was rapidly extracted for atrial myocyte isolation (see below). All animal procedures followed institutional rules (Tötungsanzeige) as reviewed by the IACUC and the Veterinarian State Authority (LAVES, Oldenburg, Germany).