In Ovo Electroporation
Electrode Recommendation and Protocol
Electrode Recommendation:
For in ovo electroporation, we recommend:
- CUY611 series or
If the target is a specific part of neural tube, we recommend a combination of:
- CUY611 series (or CUY613 series) and
- CUY614
Initially, the CUY611 series was designed for gene transfection into the neural tube. With the CUY611 series, a pair of electrodes held by an electrode holder CUY580 must be moved downwards in a long vertical line by a manipulator and will be placed on the target part of the embryo.
Some clients, however, who have to contend with a large number of samples on a daily basis find it difficult to manipulate a pair of electrodes vertically. Such clients prefer to move the pair of electrodes horizontally. The CUY613 series was designed for such clients.
Generally speaking, the CUY611 series is used for gene transfection into the ectoderm or neural tube of the chick embryo and the CUY613 series is used for gene transfection into the mesoderm and endoderm. Both series require the electrode holder CUY580.
The appropriate electrode for one’s application depends on the size of the target where the genes are to be delivered. If the length of the target is 1mm, the CUY613P1 is appropriate. However, if one would like to deliver the genes to a more specific target area, we recommend the combination of CUY613 and tungsten needle CUY614 w/CUY615. Please note the ‘Tip for in ovo electroporation’ below. It describes how a neural tube is placed between the electrodes and then the electrodes are pushed slightly against an embryo, so that the neural tube swells up. As one will find, this will increase the efficiency of gene transfection.
Protocol Recommendation:
Kindly note the file titled: ‘EP with Optic Vesicles’. This file includes the following:
- An article titled: ‘Cooperative action between L-Maf and Sox2 on d-crystallin gene expression during chick lens development’
- Drawings which demonstrate how to target various different parts of the Optic Vesicles by changing the position of an electrode and combination of electrodes, and
- An electroporation protocol:
|
Pulse Length |
Voltage |
No. of Pulses |
|
50msec |
7mV |
3 times |
Download ‘EP with Optic Vesicles’
Kindly also note the file titled: ‘In Ovo Electroporation’. This file includes a ‘tip for in ovo electroporation and various publications with relevant protocol information. The file includes the following five articles and in ovo electroporation tip:
- ‘Collinear activation of Hoxb genes during gastrulation is linked to mesoderm cell ingression’
- ‘Efficient Ectopic Gene Expression Targeting Chick Mesoderm’
- ‘Mini-Review – Misexpression of genes in brain vesicles by in ovo electroporation’
- ‘‘Shocking’ developments in chick embryology: electroporation and in ovo gene expression’
- ‘Early subdivisions in the neural plate define distinct competence for inductive signals’
- Diagrammatic tip for in ovo electroporation
Download ‘In Ovo Electroporation’
Kindly also note the file titled: ‘Heart’. It includes the following article:
- Tbx5 specifies the left/right ventricles and ventricular septum position during cardiogenesis
In Ovo Electroporation Resource:
Kindly identify the file name of interest and click on the relevant article to download it.