Cas9-VLPs targeting BFP were produced as previously described (see Table S1 for guide sequence). Cas9-VLPs were mixed with a single-stranded DNA template (IDT) in either DPBS (Thermo Fisher Scientific), Opti-MEM (Thermo Fisher Scientific), or SE/SF/SG buffer (Lonza). Unless otherwise noted, SE buffer (Lonza) with pulse code CM-150 was utilized. The mixture was electroporated using a 4D-nucleofector (Lonza) before immediately adding to 293T cells stably expressing a BFP-to-GFP reporter (Addgene plasmid #71825). A three nucleotide conversion within the BFP gene results in GFP expression. Cells were analyzed for loss of BFP (non-homologous end joining) and gain of GFP (homology-directed repair) expression after 5–7 days on a Attune NxT flow cytometer with a 96-well autosampler (Thermo Fisher Scientific).
Se sf sg buffer
Manufactured by Lonza
SE/SF/SG buffer is a laboratory product used to maintain the pH and ionic environment in various biological applications. It serves as a buffer solution to help stabilize and preserve the properties of samples or reactions during analysis or experimentation.
Automatically generated - may contain errors
Lab products found in correlation
4d nucleofector, by Lonza (2 mentions)
Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (2 mentions)
Opti mem, by Thermo Fisher Scientific (2 mentions)
Attune nxt flow cytometer, by Thermo Fisher Scientific (2 mentions)
96 well autosampler, by Thermo Fisher Scientific (2 mentions)
Se buffer, by Lonza (2 mentions)
Bfp to gfp reporter, by Addgene (2 mentions)
2 protocols using se sf sg buffer
1
Cas9-VLP-Mediated Gene Editing in BFP Cells
2
Cas9-VLP-Mediated Gene Editing in BFP Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cas9-VLPs targeting BFP were produced as previously described (see Table S1 for guide sequence). Cas9-VLPs were mixed with a single-stranded DNA template (IDT) in either DPBS (Thermo Fisher Scientific), Opti-MEM (Thermo Fisher Scientific), or SE/SF/SG buffer (Lonza). Unless otherwise noted, SE buffer (Lonza) with pulse code CM-150 was utilized. The mixture was electroporated using a 4D-nucleofector (Lonza) before immediately adding to 293T cells stably expressing a BFP-to-GFP reporter (Addgene plasmid #71825). A three nucleotide conversion within the BFP gene results in GFP expression. Cells were analyzed for loss of BFP (non-homologous end joining) and gain of GFP (homology-directed repair) expression after 5–7 days on a Attune NxT flow cytometer with a 96-well autosampler (Thermo Fisher Scientific).
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