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Digital image of 3 plasmid restriction digests run on a 1% w/v agarose gel, 3 volt/cm, stained with ethidium bromide. The DNA size marker is a commercial 1 kbp ladder. The position of the wells and direction of DNA migration is noted.
1 2 3 A 1% agarose 'slab' gel under normal light, behind a perspex UV shield. Only the marker dyes can be seen: The gel with UV illumination, the ethidium bromide stained DNA glows orange: Digital photo of the gel. Lane 1. Commercial DNA Markers (1kbplus), Lane 2. empty, Lane 3. a PCR product of just over 500 bases, Lane 4.
Gel conditions are 1% agarose, 3 volt/cm, and ethidium bromide stain. A molecular-weight size marker , also referred to as a protein ladder , DNA ladder , or RNA ladder , is a set of standards that are used to identify the approximate size of a molecule run on a gel during electrophoresis , using the principle that molecular weight is inversely ...
The most commonly used dye in agarose gel gel electrophoresis of DNA and RNA, dating as far back as the 1970s, is ethidium bromide (2,7-diamino-10-ethyl-9-phenylphenanthridiniumbromide). [ citation needed ] Ethidium Bromide (EtBr) is an orange-colored fluorescent intercalating dye.
Ethidium bromide (or homidium bromide, [2] chloride salt homidium chloride) [3] [4] is an intercalating agent commonly used as a fluorescent tag (nucleic acid stain) in molecular biology laboratories for techniques such as agarose gel electrophoresis.
The RNA samples are most commonly separated on agarose gels containing formaldehyde as a denaturing agent for the RNA to limit secondary structure. [11] [12] The gels can be stained with ethidium bromide (EtBr) and viewed under UV light to observe the quality and quantity of RNA before blotting. [11]
Agarose gels do not have a uniform pore size, but are optimal for electrophoresis of proteins that are larger than 200 kDa. [10] Agarose gel electrophoresis can also be used for the separation of DNA fragments ranging from 50 base pair to several megabases (millions of bases), [11] the largest of which require specialized apparatus. The ...
The lower the concentration of the gel, the larger the pore size, and the larger the DNA that can be sieved. However low-concentration gels (0.1 - 0.2%) are fragile and therefore hard to handle, and the electrophoresis of large DNA molecules can take several days. The limit of resolution for standard agarose gel electrophoresis is around 750 kb ...
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