Basic Steps For Digoxigenin-labeled Probes
- permeabilisation of cells with proteinase K to open cell membranes (around 25 minutes, not needed for tissue sections or some early-stage embryos)
- binding of mRNAs to marked RNA probe (usually overnight)
- antibody-phosphatase binding to RNA-probe (some hours)
- staining of antibody (e.g. with alkaline phosphatase)
The protocol takes around 2-3 days and takes some time to set up. Some companies sell robots to automate the process. As a result, large-scale screenings have been conducted in laboratories on thousands of genes. The results can usually be accessed via websites (see external links).
Read more about this topic: In Situ Hybridization
Famous quotes containing the words basic and/or steps:
“The man who is admired for the ingenuity of his larceny is almost always rediscovering some earlier form of fraud. The basic forms are all known, have all been practicised. The manners of capitalism improve. The morals may not.”
—John Kenneth Galbraith (b. 1908)
“Let there be no steps backward. A thought as to the manliness of persevering, of the want of manliness in yielding to depression, came to his rescue.”
—Anthony Trollope (1815–1882)