This phosphoramidite isused for synthesis ofoligonucleotides 5'-labeled with TAMRA
This phosphoramidite is?used for synthesis of?oligonucleotides 5'-labeled with TAMRA.
TAMRA(carboxytetramethylrhodamine) is?a?xanthene dye from the rhodamine family with emission in?the orangespectrum range (maximum at?563?nm). This fluorophore is?traditionally used as?a?FRET-acceptor(and a?quencher) in?a?pair with fluorescein (FAM) due to?significant overlapping of?theirspectra. Thus, this phosphoramidite is?convenient for the synthesis of?dual-labeled probes TaqMan, whichcontain 5'-terminal TAMRA and FAM in?the middle of?the sequence or?at?the 3'-end(using Fluorescein dT?Phosphoramidite and FAMCPG, respectively).
TAMRA 5'-labeled oligonucleotides are commonly used for quantitative PCRand fragment analysis (for example, for microsatellite marker analysis) because the equipment available has a?detectionchannel for TAMRA frequently.
The TAMRA dye is?not stable in?the presence of?ammonium andsterically non-hindered primary amines, so?it?is?strongly recommended to?follow specifiedconditions for labeled oligonucleotide deprotection.
UsageCoupling: 7.5?min.
Deprotection: tret-buthylamine : methanol : water 1 : 1 : 3?(v/v/v) (?TAMRA cocktail?) for 6?hours at?60?°C, then cool down to?room temperature.
Due to?complete and irreversible degradation of?the TAMRA dye, do?NOT use aqueous ammonium and AMA for deprotecting a?modified oligonucleotide from the solid-phase support.
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