What state should the cells be in for best results with the Neon Nxt Electoporation System?
For best results with the Neon Nxt Electoporation System, the cells should be healthy (not stressed) and actively dividing so that the nuclear membrane integrity is already weakened for payload needing to enter the nucleus. Consider “starving” the cells of serum or other essential nutrients.
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I'm experiencing arcing issues with the Neon Nxt Electoporation System. What could be causing this?
Arcing could be caused by high voltage or pulse length settings, high salt or contaminants in the DNA sample, incorrect cell density, and air bubbles formed during mixing of cell samples. Mix samples gently to avoid air bubble formation and pipette samples in a slow, smooth motion to avoid air uptake.
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After electroporation using the Neon NxT Electroporation System, how long should I wait before analyzing protein expression?
The optimal time for analysis of protein expression is dependent upon the stability of the protein being expressed. For a short-lived protein, like luciferase, analysis of protein expression should be done at 6-18 hours after electroporation. For a more stable protein, such as GFP, you should start the analysis at 24 hours or longer post-electroporation.
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How should I clean the Neon NxT Electroporation Device and the Neon NxT Pipette Station?
Clean the surface of the Neon NxT Electroporation Device and Neon NxT Pipette Station with 70% ethanol/isopropyl alcohol and wipe. Do not use harsh detergents or organic solvents to clean the units. Avoid spilling any liquid inside of the Neon NxT Pipette Station. If you accidentally spill any liquid (e.g., buffer or water) inside the Neon NxT Pipette Station, disconnect the station from the main device and wipe the station using dry laboratory paper. Invert and leave the station for 24 hours at room temperature for complete drying. Do not use an oven to dry the Neon NxT Pipette Station.
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What is the best time for analysis of CRISPR/Cas9-based KO/KI application in T cells after electroporation using the Neon NxT Electroporation System?
We recommend analyzing the cells 72 hours post-electroporation.
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