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Biotin is used in the 5' labeling of synthetic oligonucleotide probes. Biotin labeling can be detected through the use of enzyme labeled avidin or streptavidin conjugates. 5' Biotin amidite can only be added to the 5' terminus of the oligonucleotide, thus terminating the synthesis. This amidite has a DMT group which may be left on for RP cartridge purification.
Properties:
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Chemical Name:
[1-N-(4,4'-dimethoxytrityl)-biotinyl-2-aminoethoxyethyl]-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite
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Formula:
C45H62N5O7PS
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Molecular Weight:
832.04
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Appearance:
white solid
Product usage:
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Synthesis conditions:
Prior to dilution ensure that all product is at the bottom of the vial. Dilute to the recommended concentration and mix thoroughly in the sealed vial to ensure that all contents are dissolved. For optimal coupling efficiency, we suggest adding 3-8 molecular sieves per 100 µL(Aldrich # 33, 427-8; 3Å, 3.2 mm) to the amidite solution.
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Dilution:
100 µmol/mL
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Deprotection conditions:
The deprotecting conditions for this amidite will be depend on the types of amidites used for the synthesis of the oligo. If using fast deprotecting amidites deprotect in concentrated NH4OH for 1 hour at 60°C. If using standard amidites deprotect in concentrated NH4OH for 5 hours at 60°C. The DMT group may be left on or removed prior to cleavage of the oligo.
Biotin amidite is hygroscopic and may lose activity if used or stored improperly.
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The mass this product adds after conjugation and work-up (the additional mass seen by mass spectrometry) is:
392.4
Image of cleaved and deprotected structure:
Storage and handling:
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Shipping conditions:
Cold
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Storage conditions:
-15 to -30 °C
