Plasmid Transfer Efficiency
Plasmid Transfer Efficiency - Related Videos
02:30
F Plasmid Transfer and Dual Antibiotic Selection in E. coli
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2025
Source: Erdogan, F.,et al., Conjugative Mating Assays for Sequence-specific Analysis of Transfer Proteins Involved in Bacterial Conjugation. J. Vis. Exp. (2017)This video demonstrates the procedure for transferring an F plasmid from donor to recipient E. coli cells using a rescue plasmid and selecting transconjugants through dual antibiotic resistance. The process includes mixing bacterial cultures, incubation without shaking, disrupting mating pairs, and plating on selective media to confirm...
06:25
Use of In Vivo Assembly for High-efficiency Plasmid Construction
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Cited by 2 •
2025
In vivo assembly is a ligation-independent cloning method that relies on intrinsic DNA repair enzymes in bacteria to assemble DNA fragments by homologous recombination. This protocol is both time and cost-effective, as few reagents are required, and cloning efficiency can be as high as 99 %.
04:46
Changing the Direction and Orientation of Electric Field During Electric Pulses Application Improves Plasmid Gene Transfer in vitro
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Cited by 2 •
2011
Gene transfection by electroporation is improved approximately two times when orientation of electric field is changed during pulse application, while cell viability is not affected. The increase in gene transfection is caused by the increase of the membrane area which is made competent for DNA entry into the cell.
03:56
Delivery of DNA Plasmids into Urothelium of Mouse Bladder: An Electroporation-based Procedure to Efficiently Deliver DNA Plasmids into Urothelial Cells of Murine Bladder to Generate Genetically Engineered Models
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2023
This video demonstrates the delivery of DNA plasmid into the urothelium of the mouse bladder using a combination of urethral catheterization and electroporation techniques. The described procedure allows the generation of mouse bladder cancer models.
08:16
Plasmid Purification
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2023
Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under study. In this video, a step-by-step generalized procedure is described for how to perform plasmid purification.
FAQs
The delivery route affects how plasmid DNA crosses the cell envelope, so transformation, electroporation, and conjugation can produce different transfer outcomes. Comparing these approaches helps researchers identify suitable DNA delivery conditions for a particular host strain. This comparison is especially useful when an experiment requires enough modified cells for downstream cloning, protein production, or functional studies.
Selectable-marker expression provides an observable way to identify recipient cells that have acquired the introduced plasmid. Researchers can use this signal to estimate how effectively a delivery approach produced genetically modified cells. Because successful transfer also requires plasmid maintenance, marker-based assessment supports both initial selection and evaluation of whether the introduced genetic material persists.
Host strains can differ in how well they receive and maintain introduced plasmids, making strain selection an important variable in genetic experiments. Comparing strains under appropriate delivery conditions helps identify a host that yields enough modified cells for the intended application. This comparison can improve experimental reliability and guide choices for cloning, protein production, or functional studies.
Researchers assess the outcome by determining the proportion of recipient cells that successfully acquire and maintain the plasmid, often using expression of a selectable marker as evidence. The resulting measure allows different delivery methods, DNA delivery conditions, or host strains to be compared. Such comparisons reveal which setup is most effective for generating genetically modified cells.
High efficiency is valuable when an experiment needs a sufficiently large population of genetically modified cells, such as during cloning, protein production, functional studies, or genome engineering. Producing more successful recipients can improve experimental reliability while reducing the time and resources spent on selection and screening. The preferred efficiency level therefore depends on the study's downstream requirements.
In genetic engineering, transfer efficiency connects DNA delivery with the availability of cells that can be selected and used in later experiments. Measuring it helps researchers optimize delivery conditions and compare host strains before committing to larger workflows. The resulting information supports more predictable generation of modified cells for cloning, protein production, functional analysis, and genome engineering.