In utero microinjection and electroporation was performed at E14.5 essentially as described (Tabata and Nakajima, 2001 (link)), using timed pregnant CD-1 mice. Timed-pregnant mice were anesthetized and each uterus was exposed under sterile conditions. Plasmid solutions containing 1 µg/µl of DNA were injected into the lateral ventricles of the embryos using a heat-pulled capillary. Needles for injection were pulled from Wiretrol II glass capillaries (Drummond Scientific) and calibrated for 1 μl injections. DNA solutions were mixed in 10 mM Tris, pH 8.0, with 0.01% Fast Green. Forceps-type electrodes (Nepagene) with 5 mm pads were used for electroporation (five 50 ms pulses of 45 V using the ECM830 electroporation system, Harvard Apparatus). Embryos were placed back into the abdominal cavity, and mice were sutured.
Forceps type electrodes
Manufactured by Nepa Gene
Sourced in Ireland
Forceps-type electrodes are a type of laboratory equipment used for handling and manipulating small objects or samples. They have a forceps-like structure, with two opposing tips or jaws that can be used to grip and hold items securely. These electrodes are typically used in various scientific applications that require precise manipulation of delicate materials or components.
Automatically generated - may contain errors
5 protocols using forceps type electrodes
1
In Utero Microinjection and Electroporation
2
In Vivo Neuronal Electroporation Technique
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
E14.5 embryos were electroporated with 1 μg pCAG-Gfp and collected at e15.5. Injection needles were pulled from Wiretrol II glass capillaries (Drummond Scientific, Broomall, PA, USA) and calibrated for 1 μl injections. Plasmid DNAs were resuspended in 10 mM Tris, pH 8.0, with 0.01% Fast Green. Forceps-type electrodes (Nepagene, Sonidel, Dublin, Ireland) with 5 mm pads were used for electroporation (five 50 ms pulses of 25 V at E12.5 or five 50 ms pulses of 40 V at E14.5). Newborn (P0) mice were electroporated with a pcx-Gfp plasmid as previously described.8 (link)
3
In utero Jag1 overexpression in Fzd3 mutants
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Jag1 coding sequence was recovered from plasmid Addgene 17336 (ref. 58 (link)) encoding the rat Jag1 (mouse and rat Jag1 proteins are 99% identical), and cloned in the Cdk5r vector (kindly provided by Dr Paola Arlotta). The construct was verified by sequencing. The plasmid was electroporated together with a green fluorescent protein (GFP) encoding plasmid. In utero microinjection and electroporation was performed at E13.5, using pregnant Fzd3+/− females; mutant embryos were identified by their looptail phenotype. Foetuses were examined at E18.5 and genotypes were confirmed using tail clips. Needles for microinjection were pulled from Wiretrol II glass capillaries (Drummond Scientific) and calibrated for 1 μl injections. DNA solutions were mixed in 10 mM Tris, pH 8.0, with 0.01% Fast Green. Forceps-type electrodes (Nepagene) with 5 mm pads were used for electroporation (five 50-ms pulses of 30 V)59 (link).
4
In utero microinjection and electroporation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In utero microinjection and electroporation was performed at E12 as described69 (link). Briefly, timed-mated CD1 mice (Charles River Laboratories) were anesthetized and the uterine horns were exposed. 1 µl of DNA solution containing 0.5 µg/µl of pCAG-mCherry and 2 µg/µl of pCAG-Let-7 sensor or pCAG-miR1-sensor in 10 mM Tris (pH 8.0) and 0.01% Fast Green was injected per embryo. Forceps-type electrodes (Nepagene) with 5-mm pads were used for electroporation (five 50-msec pulses of 25 V). Electroporated embryos were collected 18–24 hours post-electroporation and processed for immunostaining.
5
In Utero Electroporation of Mouse Embryos
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!