piggyBac Transposon System Reagent

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Cat.No: EDY004-Y01

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Quantity

 

Description

This kit is a piggyBac Transposon System Kit, providing cells and PB system plasmids. As a non-viral vector, the piggyBac transposon system offers several advantages over traditional viral vectors: it has high safety, is easy to use (allowing direct plasmid transfection into cells), supports large vector capacity for co-expression of multiple genes, allows adjustment of helper and transposon plasmid ratios to improve integration efficiency of exogenous genes, enables precise determination of gene insertion sites by inverse PCR, achieves precise excision of the transposon upon re-transposition, has a broad host range, high transposition efficiency, and minimal reliance on host factors.

 

Kit Composition

 

Component Specification

Storage Temperature

PB System Plasmids

 Helper Plasmid (500ng/μL)

pCAG-PBase

4 μg

-20 

Transposon Plasmid500 ng/μL

PB-CMV-MCS-EFIα-CopGFP-T2A-Puro

10 μg

-20 

 

Plasmid Map Diagram

 

 

 

Note: Promoters available: CMV, CAG, EF1α

 

 

Workflow Diagram

 

 

 

Schematic Diagram of PB Gene Transposed Cell Line Construction Workflow

 

 

 

 

FAQ

Does the piggyBac Transposon System Kit recommend lipofection or electroporation?
The PB system in this kit includes plasmids compatible with lipofection, electroporation, or other transfection methods. You can select the transfection method and parameters based on cell type. Most adherent cells are compatible with various transfection methods, while electroporation is recommended for therapeutically relevant cells.
Can cells constructed with the piggyBac Transposon System Kit be stably passaged?
Yes. The piggyBac Transposon System Kit targets genomic DNA for gene transposition, allowing the genotype of target cells to be stably inherited. Note: Maintain transposed cells with half-dose puromycin for stability.
How can I verify the transposition activity of the piggyBac Transposon System Kit?
The transposon plasmid in this kit contains the CopGFP gene. Successfully transposed cells will exhibit green fluorescence under a fluorescence microscope. Due to cell heterogeneity, transfection and transposition efficiencies may vary across cell types. To minimize transposition variability, optimize transfection methods for target cells.
What should I do if the transfection efficiency in target cells is low?
Before the main experiment, conduct a transfection pre-experiment to explore different transfection methods and optimize conditions. Efficient transfection is essential for successful gene transposition.
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