What I'm doing: I am using DIG oligo probes to label mRNA in fish retina.
I am trying to adapt available protocols to what suits my needs and I'm left with a few questions...
1. What is the purpose of the dehydration step and is it an essential one - or is it more important for chromosome labeling?
2. Is the addition of salmon sperm or tRNA a necessity (some protocols include it, others use protein blocks)?
3. I've noticed that probe lengths can vary widely. Are super short probes of ~20 bases ridiculously short?
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