Whole-Mount In Situ Hybridization Protocol

RNA probes were produced with digoxigenin or fluorescein labeling mix (Roche). huc, her4, myoD, dlc, tbx24, her1 and mib plasmids were used as templates26 (link)44 (link)45 (link)46 (link)47 (link). The templates of dystrophin, foxc1a, fgf8a, papc, tcf15 and p53 were generated by RT-PCR with primers (listed in supplementary Table 6). The protocol of WISH followed the instruction of previous literatures48 (link)49 (link)50 (link). Embryos were mounted in 95% glycerol (Sigma, cat:15523)/PBST (UniRegion Bio-Tech, cat:UR-PBS001 and Tween20) pH 5.5 or flat-mounted in 95% glycerol/PBST pH 5.5 or 2:1 benzyl benzoate (Sigma, cat:B6630)/benzyl alcohol (Alfa Aesar, cat:041218). Images were captured by Zeiss Axiovision Imager A1 or Zeiss Discovery V8.

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Hsu C.H., Lin J.S., Po Lai K., Li J.W., Chan T.F., You M.S., Tse W.K, & Jiang Y.J. (2015). A new mib allele with a chromosomal deletion covering foxc1a exhibits anterior somite specification defect. Scientific Reports, 5, 10673.

Publication 2015

digoxigenin fluorescein labeling mix Axiovision Imager A1

Benzyl alcohol Benzyl benzoate Digoxigenin Dystrophin Embryos Fluorescein Glycerol Plasmids Primers Rna probes Rt pcr Tween20

Corresponding Organization :

Other organizations : National Health Research Institutes, University of Hong Kong, Chinese University of Hong Kong, Hong Kong Baptist University, National Chung Hsing University, National Taipei University

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Protocol cited in 2 other protocols

Variable analysis

independent variables

RNA probes produced with digoxigenin or fluorescein labeling mix (Roche)

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

Positive controls: Not specified
Negative controls: Not specified

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