Application Description
Biotin (BB)TM Phosphoramidites and Supports :
Biotin BBTM is a trademark of Corporation. The starting Biotin for all biotin derivatives is Biotin-D(+). Biotin BBTM contains a tert-butyl-benzoyl group at one of the sec-amino groups of Biotin. This novel feature is what makes “Biotin BBTM” (Catalog # CLP-1517) superior to other commercially available biotin products, such as “Biotin dT” and “Biotin TEG”.
Another biotin product, “Biotin Phosphoramidite” can potentially cause side reactions during oligo synthesis due to the presence of the free urea moiety of the biotin. As repeated synthesis cycles are performed, this side reaction will occur when 3’-end attachment is desired using a solid support. The “Biotin Phosphoramidite” has an additional problem of low solubility. “MMTr-Biotin Phosphoramidite”, another biotin analog has a MMTr-group on the urea portion of the biotin. The MMTr-group, however, is potentially susceptible to hydrolysis during oligo synthesis. Furthermore, the MMTr-group does not deactivate the urea moiety of biotin towards side reactions during oligo synthesis. This novel feature was originally developed by Brian Sproat and coworkers1. This study demonstrated that the protection of the urea nitrogen causes dramatic effect in the purity of the synthesized oligos.
“Biotin BBTM’’ has a 7 atom spacer utilizes this protection feature and has tert-butyl-benzoyl protection on the urea nitrogen. This product is a simple and cost effective alternative to Biotin dT.
Biotinylation within the sequence and at the 5’ Terminus:
For use in automated synthesis and associated rapid purification techniques. Applications to which our biotin phosphoramidites have been applied include:
Diagnostic probe development including occasions where multiple biotin molecules may be introduced into a probe to enhance sensitivity of ultimate detection.
Introduction of biotin in the sequence and at the 5’-terminus of a primer prior to PCR amplification and trapping of the derived biotinylated DNA using streptavidin magnetic beads.
Introduction of biotin at the 3’-terminus of a probe capable of blocking chain extension in a PCR experiment. This is achieved through Biotin-CPG.
Protection of the N-1 position of biotin with a lipophilic group induces higher solubility. It should be noted that we have found these products to be highly susceptible to deactivation by moisture in the diluent acetonitrile.
3’-End Labeling with Biotin BB CPG Solid Supports:
“Biotin BB” CPG is used for synthesis of 3’-Biotin oligomers. The tertbutyl-benzoyl protection prevents biotin chromophore unwanted side reactions. Also available are the corresponding pre-packed “Biotin BB” CPG columns listed above.
References:
1. Pieles, U.; Sproat, B.; Lamm, G. M. Nucl. Acids Res. 1990, 18, 4355.
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