Archive for the ‘DNA Transfection’ Category
Last Updated on Tuesday, 20 July 2010 08:15 Written by Administrator Sunday, 27 June 2010 08:59
DNA Transfection is the process of deliberately introducing deoxyribonucleic acid into cells. The term is used notably for non-viral methods in eukaryotic cells. It may also refer to other methods and cell types, although other terms are preferred: “transformation” is more often used to describe non-viral DNA transfer in bacteria, non-animal eukaryotic cells and plant cells – a distinctive sense of transformation refers to spontaneous genetic modifications (mutations to cancerous cells (Carcinogenesis), or under stress (UV irradiation)). “Transduction” is often used to describe virus-mediated DNA transfer. The word transfection is a blend of trans- and infection.
Genetic material (such as supercoiled plasmid DNA or siRNA constructs), or even proteins such as antibodies, may be transfected.
Transfection of animal cells typically involves opening transient pores or “holes” in the cell membrane, to allow the uptake of material. Transfection can be carried out using calcium phosphate, by electroporation, or by mixing a cationic lipid with the material to produce liposomes, which fuse with the cell membrane and deposit their cargo inside.
Transfection can result in unexpected morphologies and abnormalities in target cells.
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Last Updated on Tuesday, 20 July 2010 08:17 Written by Administrator Sunday, 27 June 2010 08:55
Optical Transfection is the process of introducing nucleic acids into cells using light. Typically, a laser is focussed to a diffraction limited spot (~1 µm diameter) using a high numerical aperture microscope objective. The plasma membrane of a cell is then exposed to this highly focussed light for a small amount of time (typically tens of milliseconds to seconds), generating a transient pore on the membrane. The generation of a photopore allows exogenous plasmid DNA, RNA, organic fluorophores, or larger objects such as semiconductor quantum nanodots to enter the cell. In this technique, one cell at a time is treated, making it particularly useful for single cell analysis.
This technique was first demonstrated in 1984 by Tsukakoshi et al., who used a frequency tripled Nd:YAG to generate stable and transient transfection of normal rat kidney cells[1]. Since this time, the optical transfection of a host of mammalian cell types has been demonstrated using a variety of laser sources, including the 405 nm continuous wave (cw) , 488 nm cw , or pulsed sources such as the 800 nm femtosecond pulsed Ti:Sapphire light.
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Last Updated on Tuesday, 20 July 2010 08:19 Written by Administrator Sunday, 27 June 2010 08:36
Electroporation is a popular method, although requiring an instrument and affecting the viability of many cell types, that also creates micro-sized holes transiently in the plasma membrane of cells under an electric discharge.
Here’s Keonwoo showing our somewhat clandestine method of electroporation. Electroporation is a way to get foreign DNA into bacteria by electrocuting them. Keonwoo needs to run because the bacteria die in a matter of seconds after being zapped. We’re the plant-bacteriology lab at Seoul National University From: www.leeskoreablog.blogspot.com
Chemical transfection and opimizationt
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Last Updated on Tuesday, 20 July 2010 08:20 Written by Administrator Sunday, 27 June 2010 08:24
DNA Transfection Efficiency
The third installment in the Big U, Big Baby series, this video is an allegory for DNA transfection.
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A short allegory for the plating of bacteria. Yousuf Khaled brings his child home, and is shocked by Margarin’s conservative attitude.
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