WISH was performed as previously described (Thisse and Thisse, 2008 (link)). Digoxigenin-labeled riboprobes for the four stag genes were synthesized from PCR clones inserted into pGEM®-T Easy vectors (Promega, Madison, Wisconsin, USA) using T7/Sp6 RNA polymerase (Roche Diagnostics, Basel, Switzerland). Anti-DIG alkaline phosphatase antibody (Roche Diagnostics, Basel, Switzerland) was used for detection, followed by visualization with nitro blue tetrazolium and 5-bromo-4-chloro-3-indolylphosphate (NBT/BCIP) (Roche Diagnostics, Basel, Switzerland). Embryos were imaged using a Leica M205 FA epifluorescence microscope (Leica, Wetzlar, Germany Applications Suite). Primers used for the amplification of stag riboprobes are listed in Supplementary Table 3 .
M205 fa epifluorescence microscope
Manufactured by Leica
Sourced in Germany
The Leica M205 FA is an epifluorescence microscope designed for high-performance imaging. It features a motorized focus drive, a large field of view, and multiple illumination options for fluorescence applications.
Automatically generated - may contain errors
2 protocols using m205 fa epifluorescence microscope
1
In Situ Hybridization of Stag Genes in Embryos
2
Whole-Mount In Situ Hybridization of Stag Genes
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WISH was performed as previously described (Thisse and Thisse, 2008) (link). Digoxigenin-labelled riboprobes for the four stag genes were synthesized from PCR clones inserted into pGEM®-T Easy vectors (Promega, Madison, Wisconsin, USA) using T7/Sp6 RNA polymerase (Roche Diagnostics, Basel, Switzerland). Anti-DIG alkaline phosphatase antibody (Roche Diagnostics, Basel, Switzerland) was used for detection, followed by visualization with nitro blue tetrazolium and 5bromo-4-chloro-3-indolylphosphate (NBT/BCIP) (Roche Diagnostics, Basel, Switzerland). Embryos were imaged using a Leica M205 FA epifluorescence microscope (Leica, Wetzlar, Germany Applications Suite). Primers used for the amplification of stag riboprobes are listed in Supplementary Table 3.
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