Solution Overview

Approach

Four Human Cell Lines were treated by the customer’s as well as industry standard Transfection Reagents (TRs) and the following characteristics compared:

1. Toxicity
2. Ability to transfect GFP
3. Ability to transfect siRNA

Brief Details:

• Human cell lines (Hela, MCF-7, HaCaT, Primary hF) were treated with increasing amounts of the TRs and the cell viability determined by MTT assay (which reveals how many cells are alive and how many dead) for each of them.
• Plasmid containing gene for GFP was used with TRs. The number of cells expressing GFP (flouroscence) was determined by microscopic observation and more precisely by FACS analysis
• siRNA against known gene (Lamin A) was transfected into the cell lines using the TRs and the efficiency determined by observing the level of silencing of the gene by western blotting and qPCR.

Key Benefits to Customer

• Based on the silencing of gene, one of the reagents was shown to be at least as effective as lipofectamine
• The analysis and data provided, enabled the customer to create the right technical and marketing brochures that helped their clients in evaluating the performance of the novel transfecting reagents over the existing ones
• Customer was able to obtain validated results with cost and time savings of close to 40 %
• Time to market for new reagent was considerably decreased