Directly labels the phosphate groups (terminal and backbone) on oligonucleotides, DNA or RNA using a TAMRA labeling reagent prepared in situ, causing less disruption of hybridization and allowing labeling that is not sequence dependent.

A number of agents have been described for labeling nucleic acids to facilitate detection of target DNA or RNA sequences. Suitable labels may provide signals detectable by fluorescence, radioactivity, colorimetry, X-ray diffraction or absorption, magnetism or enzymatic activity. It is essential that the labeling method not perturb base-pairing hybridization critical for preserving assay specificity. Nevertheless, several common methods including labeling by enzymatic incorporation can often lead to interference with the subsequent hybridization detection step, because current fluorescent labels are attached to the base (purine, pyrimidine) portion of the nucleotides where base-pairing and hybridization occurs. In addition, enzymatic labeling methods make use of additional enzymatic steps which require precise calibration to achieve a reproducible labeling yield. Moreover, because the enzymes used depend on the target type (DNA or RNA) and sequence, sequence perturbation is often observed.

To remedy this, methods of direct labeling have been used with varying degrees of success. Direct labeling through phosphodiester and phosphotriester linkages on the DNA or RNA backbone provides the additional advantage of reducing the perturbation of base-pairing hybridization. Our OliGlo™ kits utilize a direct labeling methodology through reaction with the phosphate groups (terminal and backbone) on oligonucleotides, DNA or RNA. The active labeling reagents are prepared in situ from stable precursor molecules derived from TAMRA, allowing the highly reactive labels to function at optimum efficiency for each sample.

The OliGlo™ kits allow molecular biologists and clinical researchers to label or monitor genomic DNA or RNA samples, nucleotides or oligonucleotides for easy detection and quantification. The supplied standard labeling protocol will yield labeling efficiency of approximately 10 labels per kilobase of nucleic acid depending on the purity of the sample. We found that this labeling density is sufficient for most applications. Should there be a need for adjusting labeling efficiency, the end user can simply modify the ratio of labeling dye to nucleic acid as well as incubation times for the labeling reaction as necessary. The Standard kit size provides enough material to label up to 120 μg of DNA or RNA. The trial size kit is suitable for labeling up to 35 μg of nucleic acid.

This kit is sold by Marker Gene under product code M1600/M1601.