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
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