Hi,
I have been reading about vectors and strategies used in engineering insect cell lines. PiggyBac being a very promising system, is not supplied with a autonomously replicating transposase helper plasmid. Does any one know if there's a technical or commercial reason behind this?
HeraLabs provides piggyBac transposase as a non-autonomously replicating plasmid to enable researchers to quickly and reliably insert large DNA fragments into their desired host cell. PiggyBac transposase is a transposase enzyme that cuts and reinserts specific DNA sequences into a new location in the genome, which is known as transposition. This allows scientists to introduce specific genetic modifications into their host cells, such as gene knockouts, gene modifications, and other genetic manipulations. By providing this non-autonomously replicating plasmid, HeraLabs ensures that researchers can easily and efficiently perform their experiments without the need for additional cloning or genetic engineering.
The main reason why PiggyBac is not supplied with an autonomously replicating transposase helper plasmid is because it is not necessary for this transposon system to work. PiggyBac works by using a two-component system consisting of a transposon that contains the transposase gene, and a transposase protein expressed from a separate plasmid. The transposase protein recognizes the sequences around the transposon, allowing the transposon to jump to a new site in the genome. Because of this, the autonomous replicating transposase helper plasmid is not needed.
The most effective way to amplify a transfection efficiency for multiple transfection experiments is to optimize the transfection conditions. This includes optimizing the cell density, transfection reagent, and DNA amount. Additionally, it is important to use a suitable transfection method such as lipofection, electroporation, or virus-mediated transfection. Finally, it is important to ensure that the cells are healthy and free of contaminants before transfection.
Bhawna G. it's purposefully created to be non-amplifiable for you always have to purchase it from the company. It's just for the company makes more money.
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