RNA labeling
RNA labeling is the covalent attachment of a label (such as a radioisotope, fluorescein or enzyme) to RNA for detection and detection of RNA by detecting the label. RNA labeling is widely used in the field of molecular probes and is also promising in disease diagnosis.
The ideal RNA labeling method should meet the following requirements:
1. Simple operation and high sensitivity
2. Does not affect the specificity of base pairing
3. Does not affect RNA activity
4. No effect or little effect on enzymatic activity
5. Detection method is highly sensitive and highly specific
6. Markers are harmless to the human body
RNA labeling method:
According to the reaction mechanism, the following four categories are classified: a direct labeling method, a tailing labeling method, a labeling method based on a reverse transcription reaction, and a labeling method based on RNA clonal amplification.
Methods commonly used for RNA labeling are classified according to the nature of the label:
Radioisotope labeling: 32P, 35S, 3H
Non-radioactive labeling: hapten fluorescein chemiluminescent substance
Biotin
The method of labeling RNA with biotin is generally to form biotin-UTP by binding biotin with UTP, and to synthesize labeled RNA by RNA polymerase SP6, T7 or the like by using Biotin-UTPATP, CTP and GTP as substrates.
Fluorescein
Fluorescent dyes currently commonly used include: fluorescein isothiocyanate (FITC) tetramethyl isothiocyanate rhodamine tetraethyl rhodamine Texas red algae albumin (PE) cyanine dye (Cy3, Cy5) new fluorescein Such as quantum dots (semiconductor nanocrystals).
Click chemistry
Click Chemistry is a new method for the rapid synthesis of large quantities of compounds proposed by American chemist Sharpless, winner of the 2001 Nobel Prize in Chemistry. This technology has broad application prospects in many fields, especially bio-coupling technology and biomedicine.
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