Whole-mount in situ Hybridization
Zebrafish Development
Function? Over-expressionknockdown DNA: using tissue specific promoter RNA: in vitro transcription Morpholino Dominant negative form Antagonist Whole mount in situ hybridization
Digoxigenin (DIG) DIG is a steroid hapten from Digitalis lanata ( 毛地黃 )
Structure of DIG-Modified Nucleotides Coupled via Alkali-Stable Ether Bond Formula: C 43 H 61 N 4 O 21 P 3 Li 4 Molecular Weight: DIG-UTP (R1=OH, R2=OH) DIG-dUTP (R1=OH, R2=H) DIG-ddUTP (R1= H, R2=H)
Non-Radioactive RNA-Labeling
TA vector SP6T7 Probe HindIII EcoRI HindIII
Detection methods Chemiluminescent detection CDP-star or CSPD Colorimetric detection NBT/BCIP Fluorescent detection
science.com/prodinfo_fst.htm?/DIG/dig_la beling03.htm
mRNA: FGF8 (Fibroblast Growth Factor 8) Probe: Sef (Similar Expression of Fgf gene)
(Endocrine Reviews 2005, 26: 63-77)
(Cytokine & Growth Factor Reviews 2005, 16: )
(Nature Cell Biology 2002, 4: )
(Mechanisms of Development 2002, 119: )
(Development 2002, 129: )