Product: qPCR Probes
Probes for quantitative PCR
Hylabs offers qPCR probes that are optimized for use in real-time quantitative PCR: Dual labelled probes, Double quenched probes and LC probes.
Quantitative PCR (qPCR) with probe-based methodology involves monitoring real-time fluorescence emitted by a fluorescently-labeled probe, specific to the target DNA, as it undergoes exonuclease cleavage in the 5ʹ-3ʹ direction. This allows for the measurement of DNA amplification at every cycle of the PCR process.
We offer different qPCR probes which are divided to 3 different categories: DLP / DQP / LC probes
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Dual labelled probes (DLPs) that are specifically optimized for use in real-time quantitative PCR. These DLPs are produced using advanced synthesis and purification protocols to reduce background noise to a minimum. Additionally, hylabs offers a range of reporter-quencher combinations suitable for both probe hydrolysis and hybridization-based assays, making them suitable for single or multiplex real-time PCR applications. The Dual labelled probes are offered in final yields of nmoles delivered, with a transparent and best price-performance scheme for a broad selection of Dual labelled probes.
Double quenched probes (DQPs) are optimized for use in real-time quantitative PCR and are designed to efficiently reduce background noise in probes longer than 25 nts. The length of the linear hydrolysis probe used in 5′ nuclease assays is carefully designed to balance quenching efficiency and binding stability. Sometimes it is necessary to extend the probe sequence beyond the 20-30 nt window to achieve the necessary probe Tm, especially for AT-rich target sequences. However, increasing the sequence length can diminish the quenching efficiency and result in poor signal-to-noise values. To address this, an internal quencher can be introduced closer to the fluorescent 5′ moiety to reduce background fluorescence, and an additional 3′ quencher can act as a Polymerase inhibitor to prevent the probe from being extended upon hybridization. Hylabs’ range of reporter-quencher combinations offered in their portfolio are suitable for single or multiplex real-time PCR applications and can include “non-natural” nucleotides such as LNA moieties. Like their DLPs, hylabs offers their DQPs in final yields of nmoles delivered with a transparent and best price-performance scheme for their broad selection of DQPs.
LightCycler® hybridisation probe system is based on two oligonucleotide probes that hybridize to adjacent sequences of the target DNA in a head-to-tail arrangement. Each probe is labeled with a different marker dye, which allows for FRET to occur when the probes are in close proximity to each other. hylabs offers LC® probes in various scales at defined molar amounts and synthesizes them in an ISO-certified manufacturing facility using processes licensed by Roche. The use of a spacer between the two probes is recommended to prevent steric hindrance between the dyes