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Which program should I use for co-transfection of RNP and CRISPR/Cas9 complexes ? The paper "Optimized RNP transfection for highly efficient CRISPR/Cas9-mediated gene knockout in primary T cells from Aikiko Seki and Sascha Rutz showed different programs as you have in the Lonza optimized Nucleofector Protocols?

Their publication in the Journal of Experimental Medicine is an advanced optimization for gene knock out in primary human and mouse T cells. The final protocol achieved 85-98% gene knock out for several tested receptors.The group has tested...
Missing: stable lines

The basic protocol for adherent Nucleofection« of neurons using the 4D-Nucleofector® Y Unit recommends to plate 150,000 freshly isolated or 300,000 cryopreserved neurons. What can I do if do not have enough cells to perform several samples?

Once you have found your optimal conditions you can always try to reduce the recommended cell number. To find out if lower cell numbers have been successfully used already, please check our cell data base. If you would like to reduce the cell number...
Missing: stable lines

What is the difference between the FBM™ Fibroblast Growth Basal Medium CC-3131 and the SCBM™ Stromal Cell Growth Basal Medium CC-3204? Can the FBM™ Basal Medium and the SCBM™ Basal Medium be used interchangeable? Is there a phenol red free version of the FBM™ Basal Medium?

The FBM™ Basal Medium and the SCBM™ Basal Medium are essentially the same except that the FBM™ Basal Medium contains phenol red and the SCBM™ Basal Medium does not. The FBM™ Basal Medium and the SCBM™ Basal Medium can always be used interchangeably...
Missing: stable lines

When storing my Lonza Bacterial Endotoxin Test (BET), LAL or PyroGene™ Recombinant Factor (rFC), endotoxin detection kit the temperature of my refrigerator/freezer has been out of range. Are my reagents ok to use?

...components are very stable, but you should discard any reconstituted reagents. We recommend performing an initial qualification assay to confirm that the reagents still meet the performance characteristics required by the FDA. The PyroGene rFC assay kit must...
Missing: cell lines

In my Nucleofection® Experiment, how critical is the time point for analysis post transfection?

...protein (like luciferase) analysis should be done at 6-18 hours after Nucleofection®. For a more stable protein, such as GFP, one can wait to start analysis for up to 24 hours or longer post Nucleofection®. As Nucleofection® delivers DNA directly...
Missing: cell lines

How do I handle my Lonza Control Standard Endotoxin (CSE) for Bacterial Endotoxin Testing (BET)?

Lyophilized E. coli endotoxin should be stored at 2-8°C. Endotoxin solutions are generally very stable upon storage except at more dilute concentrations. Rehydrated E. coli endotoxin should be stored refrigerated only (at 2-8°C and not frozen...
Missing: cell lines

How should I handle the reconstituted Lonza LAL PYROGENT™ Gel Clot Bacterial Endotoxin Testing (BET) Reagent?

Rehydrated LAL reagent may be stored for as long as 24 hours, but it must be kept at 2-8 °C. Do not pre-incubate lysate in the LAL test tubes before adding samples to them. This is not necessary and may cause sensitivity loss in the LAL reagent. If...
Missing: cell lines

What is the difference between a Tissue Acquisition Number (TAN) and a lot/batch number?

...Lonza will only isolate one cell type from a single piece of tissue and all product from that tissue will be processed at the same time. In such a circumstance, there would only be one TAN (for the single, initial piece of tissue) and one lot/batch...
Missing: stable lines

Can I apply more than one program (transfection condition) per plate with the 96-well Shuttle® System?

With the 96-well Shuttle® system, it is possible to apply up to 96 different programs (transfection conditions) per 96-well Nucleocuvette® Plate. Therefore, optimization of Nucleofection® Conditions for a particular cell line can...
Missing: stable

Why do plasmids that contain LTR sequences often have lower transfection efficiency?

...This is particularly true in primary cells, which have all of the normal cellular self-defense mechanisms still intact, and is true to a lesser extent with cell lines. During the process of immortalization or transformation, many cell lines lose parts their DNA...
Missing: stable
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