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DNA-purity and Nucleofection: Can low A260:A280 ratios lead to both reduced transfection efficiency and cell viability?

Yes. To check the quality of your DNA we strongly recommend determining the A260:A280 ratio. It should be ideally 1.8 - 2.0 for a good DNA preparation, but least 1.6.

How can cells be cured of mycoplasma contamination?

Where contamination has occurred, and the sample absolutely cannot be discarded, the MycoZap™ Mycoplasma Elimination Reagent has been optimized to eliminate mycoplasma with minimal toxic effects on the cells. The MycoZap™ Reagent eliminates...

Can nicked DNA lead to reduced transfection efficiency?

Yes. You should verify the integrity of your DNA on an agarose gel to see if it is degraded. Compare undigested plasmid to plasmid DNA digested with a suitable single cut restriction enzyme to linearize. Supercoiled plasmid will run faster than...

Do you have any instructions available for using a hemocytometer for cell counting?

Yes. Please click on the associated pdf file for more information.

Can I use Nucleofection to create a stable knockdown in a cancer cell line?

Yes. There are several publications available showing stable transfection of an shRNA vector: Cai S et al. (2006) Nat Genet 38:1278-88 (D10.G4.1 cells) Guo F et al. (2005) Cancer Res 65:10536-44 (K562) Hideshima T (2006) Blood 107: 4053-62 (U937)...

When I transfected my cells, error 8 appeared on the display of the Nucleofector IIN, IIS or 2b. What is the reason for that error?

Error 8 occurs when there has been an arc within the cuvette during application of the electrical program. The program was begun but was interrupted because too much current was detected. Efficiency may be lower than expected due to the incomplete...

When I transfected my cells, the message "weak" appeared on the display of the Nucleofector IIN, IIS or 2b. What does that mean and what is the reason for this error?

This message means that the device cut off the pulse before it was completed (usually this is because the conductivity of the solution is at the low end of the normal range). The pulse was applied; it is just going to be a little weaker than...