Cyprinidae: Megalamphodius Sweglesi Nematobrycon

Friday, 20 July 2012

Megalamphodius Sweglesi Nematobrycon

Primer sequences for sqt mutant constructs are available upon request. For transcription, constructs were linearized with Not I. Fluorescein or Alexa 488 labelled capped RNA was transcribed with SP6 RNA polymerase (Promega) in a reaction containing 0.5 mM rGTP, rATP and rCTP, 0.375 mM unlabelled rUTP (Roche) and 0.125 mM Chromatide Alexa 488 rUTP (Molecular Probes, Invitrogen), following the manufacturers instructions. Purified mRNA was injected into one-cell embryos, and live embryos were imaged at the four-cell stage using a Zeiss Axioplan2 upright microscope and CoolSNAP Photometrics camera (Roper Scientific). Breeding pairs of fish were pre-screened for high fertilization rates.