Stark an der Universität Stanford entwickelt. Neal Burnette und unabhängig im Labor von George R. It has been a widely used technique for over three decades. Die Western Blot-Methode wurde ursprünglich 1979 im Labor von Robert Nowinski im Fred Hutchinson Cancer Research Center in Seattle von W. In this Southern blot, it is easy to determine which cells incorporated the gene and which ones did not. Southern blot is a method commonly used in molecular biology. If the probe was labeled with radioactivity, it can expose X-ray film directly.īelow is an example of a real Southern blot used to detect the presence of a gene that was transformed into a mixed cell population. Just a reminder to those who are comparing Elenas Southern blot problem with bacterial DNA to their experiences with mammalian genomic Southerns-the genome size of mammals is about 1000 times. alkaline phosphatase or horseradish peroxidase).Ĥ) The location of the probe is revealed by incubating it with a colorless substrate that the attached enzyme converts to a colored product that can be seen or gives off light which will expose X-ray film. The left panel is an electrophoretic gel stained with ethidium bromide. The probe cannot be seen but it is either radioactive or has an enzyme bound to it (e.g. This probe will form base pairs with its complementary DNA sequence and bind to form a double-stranded DNA molecule. Test prep > MCAT > Foundation 1: Biomolecules > DNA technology. The DNA fragements retain the same pattern of separation they had on the gel.ģ) The blot is incubated with many copies of a probe which is single-stranded DNA. The DNA is denature into single strands by incubation with NaOH.Ģ) The DNA is transfered to a membrane which is a sheet of special blotting paper. Southern blotting is designed to locate a particular sequence of DNA within a complex mixture. Because there are so many different restriction fragments on the gel, it usually appears as a smear rather than discrete bands. 1) DNA (genomic or other source) is digested with a restriction enzyme and separated by gel electrophoresis, usually an agarose gel.
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