Real-Time PCR assays are prone to inhibition by various substances found in many samples (clinical, soil, plant…). Carryover of reagents used for the isolation of nucleic acids can also inhibit amplification reactions. Other causes of false-negative results include target nucleic acid degradation, sample processing errors and thermocycler malfunction. Benefits Specific - Does not interfere with target amplification - Avoid amplification of endogenous genes Sensitive - Compatible with low copy templates - Available with dark quencher for maximal signal-to-noise ratio Designed for multiplex assays - Detected in the Yakima Yellow®, VIC®, JOE channel Eurogentec’s Universal Exogenous qPCR positive Controls provide an accurate way to assess the integrity of your nucleic acid extraction and amplification assay.
IPC are labelled to Yakima Yellow, FAM or Cy5; custom labelling is available on request (custom-qPCR@eurogentec.com).
Alternatively these controls can be used as positive controls in digital PCR (dPCR) applications.
Eurogentec’s Universal Exogenous qPCR Positive Control for TaqMan® Assays is an optimized Taqman control that was designed to distinguish true target negatives from false negatives due to PCR inhibition, incorrect pipetting or cycling parameters. - The optimized control can be spiked into samples before extraction (RT-SPCx-xxx) or before qPCR Assay (RT-IPCx-xxx) without compromising amplification efficiency of the target sequence. - The optimized control doesn't match with any sequence routinely found in a lab. - The optimized control is detected using a Yakima-Yellow® (VIC® equivalent)-labeled probe and the target templates are detected using probes emiting in other channels (e.g. FAM). - Avoid amplification of endogenous genes.
Note: Alternatively, the Universal Exogenous qPCR Positive Control may be used in standardised conditions as extraction yield calibrator, template quality sensor or inter-run calibrator. A given quantity of control can be spiked into samples before extraction. A relative (directly comparing samples) or an absolute (using a dilution curve of the control) quantification is performed after extraction to normalize the extraction yields of the samples. Quantitative results of the spiked control within the template or within a reference buffer (pure water, reference template…) are compared in order to reject templates where PCR inhibition is high (low quality). Add a dilution series of the optimised control on each plate and use it to normalize PCR efficiencies between plates (also for cycler to cycler data normalization).
200 rxn kit contains: - One tube (1100 µL; white cap) containing IPC primers and YY-TAMRA probe - One tube (220 µL; red cap) containing IPC template DNA 1000 rxn kit contains: - Five tubes (1100 µL each; white cap) containing IPC primers and YY-TAMRA probe - One tube (1100 µL; yellow cap) containing IPC template DNA
• For long-term storage, the Universal Exogenous qPCR Positive Control should be kept in the dark, at -20 °C in a constant temperature freezer. • For short-term storage, the Universal Exogenous qPCR Positive Control can be kept in the dark, at 4 °C to 6°C for one month. • The 10X IPC Mix should be protected from light whenever possible to avoid degradation of the probe. • Avoid multiple freeze-thaw cycles