Capped MBSN300 mRNA was synthesized using the ~ 900 base pair N terminal sequence of Myosin Binding Subunit fused to a Glycine-Serine linker and an HA tag. A mMESSAGE mMACHINE sP6 Transcription kit (Ambion) was used to generate mRNA. Embryos were dechorionated using same procedure as for live imaging and then desiccated for 5 minutes. The embryos were covered in a mixture of 75% halocarbon 700 and 25% halocarbon 27 (Sigma). The mRNA was injected into the lateral side of the embryo approximately 2.5 hours before ventral furrow formation. The excessive injection oil was wicked off and spacer coverslips (No. 1.5) and a top coverside were added. The imaging chamber was filled with halocarbon 27 oil. Control embryos were injected with 0.1xPBS.
Halocarbon 27
Manufactured by Merck Group
Halocarbon 27 is a chlorofluorocarbon compound used in various laboratory applications. It is a colorless, non-flammable gas with a low boiling point. Halocarbon 27 is commonly employed as a refrigerant, propellant, and solvent in scientific research and industrial settings.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using halocarbon 27
1
Capped MBS mRNA Injection for Embryo Imaging
2
Embryo Mounting and Imaging Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Embryos were harvested on apple juice plates, using protocols mentioned earlier [49 ]. Staged two-hour-old embryos were dechorioned by hand by rolling them over a tape band (Scotch). Dechorioned embryos were placed on the lateral side on a mounting membrane lined with glue. The glue was prepared by submerging 10 cm of Scotch tape in 4 ml Heptane for 48 hours in a shaker at 37°C. A drop of glue was placed on the mounting membrane, gently smeared evenly, and was then allowed to air dry, before placing the dechorioned embryos. After the embryos were placed on the membrane, they were submerged in a mixture of halocarbon oil (60% Halocarbon27, 40% Halocarbon700, Sigma), and then covered with a 25 × 25 mm2 glass coverslip (Corning).
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!