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Transilluminators, a standard piece of laboratory equipment, are specifically designed to visualize nucleic acids after they have undergone gel electrophoresis.
By emitting high levels of ultraviolet light (UV) or high-intensity LED light, DNA, RNA, and macromolecules like protein are made visible. Gels that have been stained can be directly placed onto the device, and in the case of UV, the wavelength can be adjusted to fit your specific application needs.
Visualizing nucleic acids such as DNA and RNA, as well as macromolecules that include protein, is an important process in many life sciences, microbiology, and biotechnology laboratories.
Before visualization can occur, DNA must be separated, identified, or purified using a common method known as gel electrophoresis. The electrophoretic migration rate of DNA through the agarose gel can depend on the weight of the sample, as well as the concentration of gel and strength of the electric field.
This technique forms bands of DNA through separation. During or directly after the process is completed, the gel medium is stained using a fluorescent dye. This stain binds to nucleic acids, so as to mark the target DNA, RNA, or protein.
Once the gel is stained, it is placed under a light source that can fluoresce or excite the stain, making the sample visible. In many cases, the light source used is UV light. Longwave UV light has been used to cause substances to glow or fluoresce under the correct conditions due to its reactivity with organic molecules.
The key application of a transilluminator is to illuminate electrophoresed agarose or polyacrylamide gels that have been stained using a fluorescent dye. In most cases, the dye is excited using ultraviolet light because of its ability to fluoresce.
Because UV radiation can be harmful, there are some alternatives available worth considering such as LED light bulbs.
Ethidium bromide is commonly introduced into the sample because of its fluorescent nature and its ability to intercalate into the DNA based on its unique structure. As the dye fluoresces, the DNA becomes visible.
A transilluminator comes equipped with a positionable blocking window to protect from direct exposure to ultraviolet light, as well as a filter to reduce the amount of visible light inside the housing. Simple to use and designed for efficiency, these devices provide an excellent excitation light source.
Operating using high-intensity blue LED light bulbs, these devices are an alternative to UV lightboxes. Light from inside the blue light transilluminator passes through a blue filter, producing an intense signal that then passes through an amber filter, providing a large viewing surface to view agarose gel containing stained DNA bands.
The LED bulbs do not pose a health risk and provide the same level of visibility as ultraviolet light. While ultraviolet radiation may possibly sunburn or cross-link your samples without the proper care, LED bulbs ensure quality images for publication or analysis without the possible risk of damage.
Moreover, the spectrum light of LED transilluminators can effectively excite many of the common nucleic acid stains used for ultraviolet radiation. Furthermore, LED bulbs have a longer life span compared to UV bulbs.
It is worth considering which kind of dye you are using in gel electrophoresis, because this may affect your observations when using a transilluminator. A few common dyes for staining and visualizing DNA include:
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