Genecks Posted January 1, 2010 Posted January 1, 2010 (edited) Truthfully, I just wanna see the pretty colors. But on a more serious note, does anyone know where I can find a picture of green fluorescent protein or some fluorescent tag being used during southern blot analysis? The southern blot wiki page says this (bold, underline section of most relevance to thread): "The membrane is then exposed to a hybridization probe—a single DNA fragment with a specific sequence whose presence in the target DNA is to be determined. The probe DNA is labelled so that it can be detected, usually by incorporating radioactivity or tagging the molecule with a fluorescent or chromogenic dye. In some cases, the hybridization probe may be made from RNA, rather than DNA. To ensure the specificity of the binding of the probe to the sample DNA, most common hybridization methods use salmon or herring sperm DNA for blocking of the membrane surface and target DNA, deionized formamide, and detergents such as SDS to reduce non-specific binding of the probe." - http://en.wikipedia.org/wiki/Southern_blot I tried looking, but I really couldn't find anything... I see plenty of autoradiograms, but I haven't really come across something that involves GFP. Maybe this is fluorescently tagged? It's from the VNTR wiki (http://en.wikipedia.org/wiki/VNTR): If anyone knows of an article or some image, I'd appreciate it. It doesn't really have to be southern blot (north and west are fine), but I'd appreciate if it were. Edited January 1, 2010 by Genecks
CharonY Posted January 1, 2010 Posted January 1, 2010 There is little reason to use GFP in Southerns. But there are uses for GFP antibodies. Search for that term, maybe in conjunction with Western and you should find something.
Genecks Posted January 2, 2010 Author Posted January 2, 2010 My mind ascended to a new level after a night of sleep and considered that GFP is a protein, and probably would not be easily used in southern blot analysis unless the GFP has a nice complementary nucleotide sequence to adhere to the nucleotide sequence it will attach to. Furthermore, it's molecular structure might actually cause it to have difficulty with adhering if it did have that nucleotide sequence (I'm not sure about that). However, since my main interest is usage of GFP or something similar, then I suspect a Western blot, as you say, may be of more interest. Thanks, CharonY.
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