LightCycler^® Probes

LightCycler probes are highly sensitive and sequence-specific fluorescent probes designed for use with the Roche LightCycler instruments.

• When to Choose LightCycler Probes
• Benefits of Using LightCycler Probes
• Add LNA^® to Your Probe
• How LightCycler Probes Work
• Product Features
• Guaranteed Yields
• Labels and Modifications for LightCycler Probes

Choose LightCycler Probes for:

• SNP detection
• Allelic discrimination
• End point detection

Several fluorophores are available and are suitable for multiplex analysis.

Benefits of Using LightCycler Probes include:

• Increased specificity
• Probe preserved during the reaction
• Increased thermal stability and hybridization specificity
• Greater accuracy in gene quantitation and allelic discrimination
• Easier and more sensitive probe designs for problematic target sequences

Add LNA to Your Probe for:

• Increased thermal stability and hybridization specificity
• Greater accuracy in gene quantitation and allelic discrimination
• Easier and more sensitive probe designs for problematic target sequences

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How LightCycler Probes Work

A LightCycler probe system consists of a pair of single-stranded fluorescent-labeled oligonucleotides. Oligo Probe 1 is labeled at the 3' end with a donor fluorophore dye and Oligo Probe 2 is labeled at its 5' end with one of two available acceptor fluorophore dyes. The free 3' hydroxyl group of Probe 2 must be blocked with a phosphate group (P) to prevent DNA polymerase extension. There should be a spacer of 1 to 5 nt to separate the two probes from each other. The structures and mechanism of a LightCycler probe are shown below.

During the annealing step of real-time quantitative PCR, the PCR primers and the LightCycler probes hybridize to their specific target regions bringing the donor dye into close proximity to the acceptor dye. When the donor dye is excited by light from the LightCycler instrument, energy is transferred by Fluorescence Resonance Energy Transfer (FRET) from the donor to the acceptor dye. The acceptor fluorophore's emission wavelength is detected. The increase in fluorescence signal is directly proportional to the amount of target DNA.

Product Features include:

• Available in lengths of 15 to 40 mers (optimal length: 20 to 30 mers)
• Simple pricing – HPLC purification and base charges are included
• Stringent quality control including electrospray mass spectroscopy
• Shipped within 7–10 business days
• Shipped in an amber tube to protect dye integrity
• Custom formats available (normalization, special plates, etc.)

Sigma's probes are provided in a format to simplify your experimental planning.

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Please inquire for alternative quantities.

The recommended constructs for LightCycler Probe 1 and LightCycler Probe 2 are listed in the tables below:

Sigma^® is a licensed supplier of a variety of dyes and quenchers and continually adds to its portfolio of new chemistries. For assistance in the design of your LightCycler probes and assays, submit your request at sigma.com/designmyprobe.

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