Dual Labeled Probe: 5' FAM Fluorophore Reporter (Emission 520 nm) 3' BHQ-1 (Black Hole Quencher)

Product Description

Dual-labeled BHQ™ Probes are traditional, linear, FRET probes (TaqMan Probes) incorporating a fluorophore and quencher covalently attached to the 5' and 3' ends of an oligo typically 20 to 30 bases in length. Fluorescence is released through the 5' exonuclease activity of Taq polymerase, which cleaves off the fluorescent dye upon the probe's hybridization to its complementary sequence. Dual-labeled BHQ Probes are ideal for detecting the presence and quantifying the amount of specific target sequences.

Design Recommendations for Dual-labeled BHQ Probes

Please be aware that sequences longer than 30 bases are unlikely to have efficient quenching as an end-labeled probe. For such situations please consider positioning the quencher internally instead.

Conversely, sequences shorter than 20 bases may have been designed as a Minor Groove Binder (MGB™) probe and are unlikely to bind at the proper temperature of 70 °C. Please consider ordering such a sequence as a BHQplus™ probe instead, or else contact us with any questions regarding probe design.

NOTE:

• Building probes with an Internal Quencher (Currently available with FAM only)
• Adding an internal modification annotated by T(...) indicates that a Thymidine base is attached to the internal modification. Please be aware that an additional T base will be introduced into the sequence when adding such an internal modification.
• Building probes with wobble/degenerate bases (What are Wobbles?)
• To ensure the most efficient processing of your oligo, use IUPAC code when entering wobble/degenerate bases in the Sequence Entry field instead of using parentheses, e.g. (A/G), (A/G/C/T), etc.

Fluorophore and BHQ dye selection chart

This chart is intended to guide you through the dye selection process for your oligonucleotide. A fluorophore and quencher combination may be selected for applications such as probe-based qPCR and SNP genotyping.