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Furthermore, the HotStarTaq polymerase present in the QuantiTect Multiplex PCR buffer in the PCR master mix is inactive at 30°C.

The Genotyping Master Mix gave the highest Cq values combined with the highest ΔRn values, indicating that this master mix is not ideal for multiplex qMSP (Table 1).

reverse primer concentrations have to be increased for use in one-step analyses due to first-strand synthesis requirements and, 3). a different master mix is employed for one-step as opposed to two-step qPCR.

Nonetheless dimers still appeared in some reactions when DNA concentration was low (i.e. below 100 copies), more often for Lux™ chemistry than for Plexor™, as Plexor™ master mix is optimized to minimize dimer formation.

For purification of a full 96-well plate of samples, a master mix of 1100 μL Exonuclease I and 550 μL recombinant Shrimp Alkaline Phosphotase is prepared, and 15 μL of the master mix is manually pipetted to each sample PCR product.

For our applications, an amplification master mix is prepared in a 15 mL conical tube using 850 μL GeneAmp® 10X PCR Buffer I (Life Technologies, Applied Biosystems), 680 μL GeneAmp® dNTP blend 10 mM (Life Technologies, Applied Biosystems), 340 μL of each 10 μM amplification primer, and 5695 μL deionized water.

5  μL of SYBR Green master mix was mixed with 1  μL of primer mix and 4  μL of diluted cDNA (1 : 10) (total reaction volume: 10  μL).

Briefly, ×2 RT master mix was prepared per kit instructions.

To minimize the chance of error, a master mix was prepared before class so students only had two pipetting steps (mixing their DNA to the PCR master mix).

Probes and Universal PCR Master Mix were obtained from Applied Biosystems.

For PHD3, TaqMan universal master mix was used.