Multiplex Probe qPCR Mix Plus U

Multiplex Probe qPCR Mix Plus U is a 2X concentrated premix for real-time quantitative PCR with probe method. In use, just add the DNA template, primer and probe to react. This product contains antibody technology modified Hotstart Taq DNA polymerase. Combined with GDSBio's special real-time PCR Buffer, it can not only effectively inhibit primers dimer and other non-specific amplification, but also improve the amplification efficiency, allowing multi-probe qPCR reaction. This reagent introduced dUTP/UDG anti-contamination system, which can remove PCR products containing dUTP before PCR reaction, effectively avoid the influence of cross contamination of amplification products on quantification. This product is convenient for use with TaqMan, primers to achieve a fully premixed Master Mix molecular diagnostic kit, and is perfectly compatible with common quantitative PCR instruments, such as ABI, Roche, Bio-Rad, etc. This product can be used with TaqMan and other fluorescent probes, and is perfectly compatible with common quantitative PCR instruments, such as ABI, Roche, Bio-rad, etc.

The reaction system of this product can be prepared at room temperature without an ice box. The prepared PCR reaction system can be placed at room temperature for 24 hours and the amplification efficiency remains unchanged.

Application
Probe gene expression analysis
Multiple Low-copy gene detection
Probe microarray validation
Probe gene knockdown validation

Features
This kit is suitable for Multiplex fluorescence quantification by probe method
This kit is compatible with many real-time systems
Hot-start technology brings high specificity and reproducible amplification
dUTP/UDG system, effectively prevent PCR product contamination

Fast Amplification Mode Supported: Run Time <50 min, Controlled ∆Ct Loss.

OEM available

Desktop Stability Test:
Using human genomic DNA (0.1ng/µl) as a template, gene detection is performed through Multiplex Probe qPCR Mix Plus U. Figures 1 and 2 are the amplification curves detected after the reaction system is prepared (0 hr) and after being placed at room temperature (25°C) for 48 hours (48 hr), respectively; at the same time, a comparative test is conducted with the manufacturer T's similar product, with Figures 3 and 4 being the amplification curves of manufacturer T's assembly completed at 0 hr and 48 hr, respectively. It can be seen that after the pre-assembled system is placed at room temperature for 48 hours, Multiplex Probe qPCR Mix Plus U still has excellent amplification performance.

High-Concentration Contamination Clearance Test (Multiplex qPCR System):
In a triplex (3-Plex) fluorescence amplification system, U-containing amplicons were deliberately introduced at 10⁴-fold dilution (concentration 100a), 10⁵-fold dilution (concentration 10a), and 10⁶-fold dilution (concentration a) to simulate laboratory carryover contamination. The anti-contamination performance of P2601 (without UDG/dUTP system) was compared head-to-head against P2701 (this product). A no-template control (NTC) was run in parallel.

Results:
P2601 (Control group — no anti-contamination system): The NTC showed no amplification. Contaminated channels (GAPDH / RPLPO / GUSB) produced strong amplification signals across all dilution gradients, indicating severe false-positive results.
P2701 (Test group — this product): Following 37°C digestion for 2 min, false-positive signals in contaminated channels were effectively eliminated (below the limit of detection), with amplification curves restored to baseline levels. The Ct value for RPLPO detection was significantly delayed (ΔCt~11), demonstrating a high clearance efficiency of up to 99.95%.The NTC channel showed no amplification.

Rapid Amplification Test:

Multiplex Probe qPCR Mix Plus U supports rapid amplification mode, with a reaction time of <50 minutes, saving >30% of time, and with controllable Ct loss: approximately 1–3.5 cycles.

Target Allocation Strategy (Recommendation): In multiplex PCR assays, optimize channel assignment to offset fast-cycling kinetic losses:

ROX Channel: Allocate high-abundance, highly expressed, or non-LoD (Limit of Detection) target genes.

FAM or CY5 Channels: Reserve for fragile, ultra-low copy targets. These channels remain virtually unaffected by fast protocols ( ∆Ct ~1).