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Can Lonza's antibiotics be used in clinical trials?

No. They are for research purposes only.

Are your Nucleofector Solutions checked for RNase activity during your QC process?

Yes. We do test the Solutions with a real time RT-PCR based assay. This assay basically measures the quantity of an RNA template after incubation in our solution. In case there is RNAse contamination the template would be digested which would be...

What kind of buffer do you recommend for resuspending my siRNA?

Please use the buffer that is recommended by your siRNA manufacturer.

What stock concentration of Propidium Iodide does Lonza use for FACS analysis?

We use a stock concentration of 100 µg/ml in PBS. This is diluted 1:300 – 1:400 into the sample.

Do you have any siRNA Nucleofection results using concentrations lower than 50nM?

Yes. Please click on the attached Technical Reference Guide: "Designing an RNAi Experiment Using Nucleofection™". On page 3 you can find a table with some examples.

The animal facility which isolates my hepatocytes is at a different location, should I be concerned about transport time?

Isolated hepatocytes can be transported in Krebs-Henseleit Buffer or supplemented William's E Medium. Long transport time for isolated hepatocytes, (in excess of one hour) can cause a decrease in efficiency of Nucleofection™, but does not seem to...

How much DNA can I use when transfecting hepatocytes with the standard Nucleofector?

We have used up to 6 µg of DNA per 100 µl reaction with no deleterious effect.

For hepatocytes, is plating density a concern post Nucleofection?

Yes, the optimized protocols for the standard Nucleofector™ recommend the use of 6 well plates. Plating into a 12-well plate will pose a problem as less cells will attach at high densities.

Four hours after Nucleofection, I can see the hepatocytes have attached to the well, but the morphology does not look correct, should I be concerned?

There is no reason for alarm. The hepatocytes may not exhibit normal morphology a few hours after Nucleofection™ but by 24 hours post Nucleofection™, morphology should be normal. Remember to perform a fluid change about 4 hours post Nucleofection™....

What are the contents of a 96-well Shuttle Kit?

As the system will most likely be used for high-throughput approaches we provide plates which are pre-equipped with 96-well Nucleocuvette™ modules (2x8). In addition to that, the 96-well Shuttle solution, the supplement, and the pmaxGFP™ are also...
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