Lonza 4D Nucleofector with Shuttle unit (V4SC-2960 Nucleocuvette Strips) was used for electroporation, following the manufacturer’s instructions. Jurkat cells were electroporated using the SF Cell Line Nucleofector X Kit (Lonza), CA-137 program, with 2 × 105 cells in 20 µL SF buffer for each nucleofection reaction. The cell suspension was mixed with RNPs, immediately transferred to the nucleocuvette, and subjected to nucleofection in the 96-well Shuttle device. Cells were immediately re-suspended in the cultivation medium and plated on 96-well, flat-bottom, non-cell culture treated plates (Falcon). Cells were harvested 48-h post-transfection for genomic DNA extraction and viability assays. For the Homology-Directed Repair efficiency assay , the HDR template, 160 nt long ssDNA (Supplementary Table 2), was collected via pipetting from the HDR plate after the RNPs addition and immediately before the electroporation. The electroporation parameters, cells recovery and proliferation were performed the same way as described above.
4d nucleofector with shuttle unit v4sc 2960 nucleocuvette strips
Manufactured by Lonza
The 4D Nucleofector with Shuttle unit (V4SC-2960 Nucleocuvette Strips) is a laboratory equipment designed for efficient nucleic acid delivery into cells. It provides a controlled environment for electroporation-based transfection, enabling the introduction of various molecules such as DNA, RNA, or proteins into cells. The device includes the 4D Nucleofector and the Shuttle unit, which work together to facilitate the transfection process. The Nucleocuvette Strips are compatible with the system and provide a convenient format for sample processing.
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2 protocols using 4d nucleofector with shuttle unit v4sc 2960 nucleocuvette strips
1
Efficient Jurkat Cells Electroporation
2
Efficient CRISPR-Cas9 Genome Editing via Nucleofection
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A Lonza 4D-Nucleofector with
Shuttle unit (V4SC-2960 Nucleocuvette Strips) was used for transfection,
following the manufacturer’s instructions. For transfection,
cells were harvested by centrifugation (200 × g, RT, 5 min) and
re-suspended at 10 × 106 cells mL–1 (2 × 105 cells 20 μL–1)
in the SF Cell Line Nucleofector X Kit buffer (Lonza), unless stated
otherwise. The cell suspension was mixed with the RNPs, immediately
transferred to the nucleocuvette, and transfected using the CA-137
Nucleofector program, except where indicated otherwise. After transfection,
the cells were immediately re-suspended in the pre-warmed cultivation
medium and plated onto 96-well, flat-bottom, non-treated plates (Falcon)
and cultured at 37 °C in 5% CO2 incubators and maintained
at a density of 0.5–1.0 × 106 cells mL–1. After 48 h, the cells were harvested for the viability
assay and genomic DNA, as described below. For the HDR template insertion,
the HDR template was added to the cells and the suspension transferred
to the RNPs immediately before transfection. The transfection parameters,
cell recovery step, and proliferation conditions were as described
above. The cells were harvested 48 h post-transfection for the viability
assessment and after 7, 14, and 21 days for GFP insertion efficiency.
Shuttle unit (V4SC-2960 Nucleocuvette Strips) was used for transfection,
following the manufacturer’s instructions. For transfection,
cells were harvested by centrifugation (200 × g, RT, 5 min) and
re-suspended at 10 × 106 cells mL–1 (2 × 105 cells 20 μL–1)
in the SF Cell Line Nucleofector X Kit buffer (Lonza), unless stated
otherwise. The cell suspension was mixed with the RNPs, immediately
transferred to the nucleocuvette, and transfected using the CA-137
Nucleofector program, except where indicated otherwise. After transfection,
the cells were immediately re-suspended in the pre-warmed cultivation
medium and plated onto 96-well, flat-bottom, non-treated plates (Falcon)
and cultured at 37 °C in 5% CO2 incubators and maintained
at a density of 0.5–1.0 × 106 cells mL–1. After 48 h, the cells were harvested for the viability
assay and genomic DNA, as described below. For the HDR template insertion,
the HDR template was added to the cells and the suspension transferred
to the RNPs immediately before transfection. The transfection parameters,
cell recovery step, and proliferation conditions were as described
above. The cells were harvested 48 h post-transfection for the viability
assessment and after 7, 14, and 21 days for GFP insertion efficiency.
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