Efficient RNA Labeling with abberior® STAR dyes

Abberior® STAR 580 and STAR RED dyes are hydrophilic fluorescent dyes with remarkable photostability that give optimal performance of dye-conjugates in STED and confocal microscopy.

Jena Bioscience now offers STAR 580- and STAR RED-labeled single nucleotides as well as the corresponding HighYield T7 RNA Labeling Kits for in vitro transcription-based direct labeling of RNA probes.

Linker-optimized abberior® STAR dye-labeled UTPs in combination with an optimized enzyme mix allow efficient preparation of abberior® STAR dye-labeled RNA probes (> 100 nt up to several thousand nt) (Fig. 1, Tab. 1).

Table 1: Single nucleotides and HighYield T7 RNA Labeling kits for efficient abberior® STAR 580 and STAR RED labeling.

Product Dye Substitute for Excmax / Emmax [nm] Recommended range of substitution
UTP-PEG5-STAR 580 abberior® STAR 580 AF555 (Alexa Fluor®555) / Cy3 / AF 594 Alexa Fluor® 594) 587 / 607 30-50%
HighYield T7 STAR 580 RNA Labeling Kit
UTP-PEG5-STAR RED abberior® STAR RED AF647 (Alexa Fluor®647), Cy5 638 / 655 20-40%
HighYield T7 STAR RED RNA Labeling Kit

Enzymatic properties of abberior® STAR-labeled UTPs are comparable to Alexa Fluor® dye-labeled counterparts:

Figure 1: In vitro transcription-based direct labeling of RNA probes with abberior® STAR dye-labeled UTPs. A) Comparison of UTP-PEG5-STAR 580 & UTP-PEG5-AF555. B) Comparison of UTP-PEG5-STAR RED & UTP-PEG5-AF647.

1.4 kbp RNA was generated using HighYield T7 AFXXX RNA Labeling Kit components in combination with an abberior® STAR or AF (Alexa Fluor®) – labeled UTP. Reaction conditions: 1 µg DNA template, 30 min at 37 °C. Crude in vitro reaction mix was subsequently analyzed be agarose gel electrophoresis (Visualization of total RNA with RNA intercalator EtBr / 0 – 100 %: Increasing percentage of UTP substitution by labeled UTP).


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