Riboprobe: Difference between revisions
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== Riboprobe == | |||
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File:Principles_of_fluorescence_in_situ_hybridization.jpg|Principles of fluorescence in situ hybridization | |||
File:Spectral_karyotyping_and_multicolor-FISH_paint_each_human_chromosome_in_one_of_24_colors.jpg|Spectral karyotyping and multicolor-FISH paint each human chromosome in one of 24 colors | |||
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Revision as of 21:13, 23 February 2025
Riboprobe is a type of RNA probe that is synthesized in vitro by RNA transcription. It is used in molecular biology research, particularly in the field of genetics, to detect the presence of complementary nucleic acid sequences (DNA or RNA) in a sample. Riboprobes are often labeled with radioactive or non-radioactive tags to enable detection.
Synthesis
Riboprobes are synthesized from a DNA template by in vitro transcription. The DNA template is usually a plasmid or a PCR product that contains the sequence of interest. The transcription is carried out by RNA polymerase, which synthesizes the riboprobe in the 5' to 3' direction. The riboprobe can be labeled during synthesis by incorporating labeled nucleotides into the RNA.
Applications
Riboprobes are used in a variety of applications in molecular biology. They are often used in Northern blotting to detect specific RNA molecules in a sample. They can also be used in in situ hybridization to detect specific sequences of DNA or RNA in cells or tissues. In addition, riboprobes can be used in RNAse protection assays to measure the amount of specific RNA molecules in a sample.
Advantages and Disadvantages
One of the main advantages of riboprobes is their high specificity. Because they are complementary to the sequence of interest, they can specifically bind to that sequence and not to other sequences. This makes them very useful for detecting specific sequences in complex samples.
However, riboprobes also have some disadvantages. One of the main disadvantages is that they can be degraded by RNases, which are enzymes that degrade RNA. This can make it difficult to store and handle riboprobes. In addition, the synthesis of riboprobes can be time-consuming and requires specialized equipment and expertise.
