ABSTRACT
The orientation of the probe nucleotide sequence must be anti-sense to that of the target.
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The label must be chosen according to the visualization method used (
see
Chapter 5). The probe structure is of one of the following types:
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Double-stranded DNA
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Generally cDNA
•
Single-stranded DNA
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Generally obtained by PCR
•
Synthetic DNA
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Oligonucleotide
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RNA
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Generally cRNA and the target nucleic acid sequences include:
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DNA in interphase nuclei
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Chromosomal DNA
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Ribosomal RNA
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Transfer RNA
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Messenger RNA
•
Viral RNA
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Principle of probe hybridization against target nucleic acid:
•
The target is oriented 3'
•
The probe is oriented 5'
3', i.e., anti-sense
Four types of probes can be defined, according to the type of nucleic acid (DNA or RNA) and its method of generation.
