16 Whole Mount in Situ Hybridization for mRNA Detection in Chick Embryos
223
•
I)AxotiOll
•
Tab:otf'be solu"oo Addlle>O/solulioo •
~ '"
.,
,...
Rock ,bedHh
4)Rthydndon
SJProoeinas< K ....",.,.",
6)Refiution
IO)Prdoybriruudon
II)Hybriruzarion
7)Heu rtealmenl
(opdolUl)
12)Tab: offproba
(Wllbaenlle
14)Bloc:ting
LJl
(In ,be dolt)
18JCOIor ....... on
• •
IS)An,lbocIyomotion
•
16)Wosh
19)DutoIninl
20JCle:Jrinl
Fig. l. Protocol of a whole-mount in itu hybridization
Materials
•
•
..
3rrmrur cr (0 iI 24--welJ dish l
BJChill on Ice
~
. , 13)Wasb
17)!;qulbrizorion In
AP buffer
21lObsemuioo
Dissection microscope (e.g., Leica MZ series, Zeiss Stemi Equipment
SVll)
- Agarose gel electrophoresis apparatus (RNase-free)
- Variable speed shaking platform (e.g., Belly Dancer)
- Heater blocks that can accommodate 1.5-2 ml microtubes
- Hybridization oven
223
•
I)AxotiOll
•
Tab:otf'be solu"oo Addlle>O/solulioo •
~ '"
.,
,...
Rock ,bedHh
4)Rthydndon
SJProoeinas< K ....",.,.",
6)Refiution
IO)Prdoybriruudon
II)Hybriruzarion
7)Heu rtealmenl
(opdolUl)
12)Tab: offproba
(Wllbaenlle
14)Bloc:ting
LJl
(In ,be dolt)
18JCOIor ....... on
• •
IS)An,lbocIyomotion
•
16)Wosh
19)DutoIninl
20JCle:Jrinl
Fig. l. Protocol of a whole-mount in itu hybridization
Materials
•
•
..
3rrmrur cr (0 iI 24--welJ dish l
BJChill on Ice
~
. , 13)Wasb
17)!;qulbrizorion In
AP buffer
21lObsemuioo
Dissection microscope (e.g., Leica MZ series, Zeiss Stemi Equipment
SVll)
- Agarose gel electrophoresis apparatus (RNase-free)
- Variable speed shaking platform (e.g., Belly Dancer)
- Heater blocks that can accommodate 1.5-2 ml microtubes
- Hybridization oven