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Commit 5daa9736 authored by Timofej Paramonov Bliki's avatar Timofej Paramonov Bliki :heart:
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Would be nice to add buttons for the protocols at the bottom, but due to time constraints I will leave this be as it's not a necessary feature.
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We wanted to amplify eGFP DNA, which would serve as control mRNA for the lipid encapsulation. We wanted to amplify eGFP DNA, which would serve as control mRNA for the lipid encapsulation.
The eGFP DNA was amplified by PCR using the following protocol: The eGFP DNA was amplified by PCR using the following protocol:
<a href="https://static.igem.wiki/teams/5342/documents/protocols/pcr-procedure-radboud-university-team.pdf" target="_blank">“PCR procedure Radboud University team”</a> and <a href="https://static.igem.wiki/teams/5342/documents/protocols/pcr-procedure-radboud-university-team.pdf" target="_blank">“PCR procedure Radboud University team”</a> and
PUC-18 GFP template. <!-- Add me! --> <a href="https://www.addgene.org/220495/" target="_blank">PUC-18 GFP template.</a> <!-- Add me! -->
After a sufficient amount of GFP DNA was received, we continued with the purification of the DNA. After a sufficient amount of GFP DNA was received, we continued with the purification of the DNA.
For the PCR purification, we used For the PCR purification, we used
<a href="https://www.qiagen.com/us/products/discovery-and-translational-research/dna-rna-purification/dna-purification/dna-clean-up/qiaquick-pcr-purification-kit" target="_blank">QIAquick PCR Purification Kit</a> <a href="https://www.qiagen.com/us/products/discovery-and-translational-research/dna-rna-purification/dna-purification/dna-clean-up/qiaquick-pcr-purification-kit" target="_blank">QIAquick PCR Purification Kit</a>
...@@ -126,7 +126,7 @@ ...@@ -126,7 +126,7 @@
This is why we performed experiments using Hek 293 cell lines. This is why we performed experiments using Hek 293 cell lines.
We encapsulated the mRNA into our LNPs and transfected the HEK cells using the lipotransfection method with the Jet messenger transfection reagent. We encapsulated the mRNA into our LNPs and transfected the HEK cells using the lipotransfection method with the Jet messenger transfection reagent.
For the GFP mRNA transfection, we used the following protocol: For the GFP mRNA transfection, we used the following protocol:
<a href="https://static.igem.wiki/teams/5342/documents/protocols/polyplus-short-protocol-jetmessenger-mrna.pdf">“Polyplus Short-protocol- jetMESSENGER-mRNA”</a> <a href="https://static.igem.wiki/teams/5342/documents/protocols/polyplus-short-protocol-jetmessenger-mrna.pdf" target="_blank">“Polyplus Short-protocol- jetMESSENGER-mRNA”</a>
and incubated the cells overnight. and incubated the cells overnight.
After the incubation, the cells were evaluated for any GFP production using the EVOS M5000 fluorescence microscope. After the incubation, the cells were evaluated for any GFP production using the EVOS M5000 fluorescence microscope.
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