What technique would you use for each senario(1 and 2) and why. FISH, immunofluorescence, and GFP fusion. proteins. Each technique is only used once.
What technique would you use for each senario(1 and 2) and why. FISH, immunofluorescence, and GFP fusion. proteins. Each technique is only used once.
1.You are studying cells in the lab from four patients with I-cell disease. First, you need to know if
the phosphotransferase gene is being expressed in any of these 4 patient’s cells.
2.Next, you want to determine if you can replace the defective protein with a normal one. First,
you need to determine if the replacement protein (one that you are creating in the lab and
inserting it into the patent’s cells), will be targeted to the lysosome.
Introduction-FISH
In this technique, the full set of chromosomes from an individual is affixed to a glass slide and then exposed to a “probe”—a small piece of purified DNA tagged with a fluorescent dye.
Principle =The basic principle involved is hybridization of nuclear DNA of either interphase cells or of metaphase chromosomes affixed to a microscopic slide, with a nucleic acid probe. The probes are either labeled indirectly with a hapten or directly through incorporation of a fluorophore
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