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Gene Expression Assay and Data Analysis
What happens if the hybridization time is extended beyond 16 hours?
The standard gene expression hybridization time is 14-16 hours at 45° C. At high temperatures and longer incubation times the sample will evaporate. Loss of sample is undesirable for several reasons:
- Low volume of hybridization solution in the probe array can lead to dry spots that will show up as uneven hybridization and thus, compromise data.
- Sample loss compromises the possibility of repeating the experiment with the identical sample.
- Sample evaporation can lead to changes in the salt concentration of the solution which can affect the stringency conditions for hybridization.