Synthesis and In Situ Hybridization of fgfr3 Probe

The template for fgfr3 antisense probe synthesis was amplified with the following primer, forward primer: 5'-GGTGACCTTGGAAAGGATTACTG-3', reverse primer: 5'-CCTGCCTCGTCCTCATCTTCATC-3'. Primers for other probes are available from the corresponding authors upon request. PCR products were cloned into the pGEMT-easy vector (Promega). Digoxigenin-labelled probes were generated by in vitro transcription (DIG RNA Labeling Kit, Roche) and in situ hybridization was carried out as previously described 30 (link). The WISH images were captured using a SteREO Discovery 20 microscope (Carl Zeiss).

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Sun X., Zhang R., Chen H., Du X., Chen S., Huang J., Liu M., Xu M., Luo F., Jin M., Su N., Qi H., Yang J., Tan Q., Zhang D., Ni Z., Liang S., Zhang B., Chen D., Zhang X., Luo L., Chen L, & Xie Y. (2020). Fgfr3 mutation disrupts chondrogenesis and bone ossification in zebrafish model mimicking CATSHL syndrome partially via enhanced Wnt/β-catenin signaling. Theranostics, 10(16), 7111-7130.

Publication 2020

pGEMT-easy vector DIG RNA Labeling Kit SteREO Discovery 20 microscope

Digoxigenin Fgfr3 In situ hybridization Microscope Primers Promega Synthesis Transcription Vector

Corresponding Organization :

Other organizations : Daping Hospital, Army Medical University, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Columbia University, Southwest University

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Variable analysis

independent variables

Forward primer: 5'-GGTGACCTTGGAAAGGATTACTG-3'
Reverse primer: 5'-CCTGCCTCGTCCTCATCTTCATC-3'

dependent variables

Expression of fgfr3 gene

control variables

PGEMT-easy vector (Promega)
Digoxigenin-labelled probes generated by in vitro transcription (DIG RNA Labeling Kit, Roche)
In situ hybridization protocol as previously described 30 (link)

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