Effective UNG digestion.
Effective UNG digestion.
Two-step RT-PCR.
Two-step RT-PCR.
Highly specific amplification.
Highly specific amplification.
Wide dynamic range in two-step PCR.
Wide dynamic range in two-step PCR.
Wide dynamic range in real-time PCR.
Wide dynamic range in real-time PCR.
High sensitivity and efficiency, and wide dynamic range.
High sensitivity and efficiency, and wide dynamic range.
Effective UNG digestion. 106 copies of two dUMP-containing PCR amplicons were treated with or without UNG and then amplified by real-time PCR. UNG was from various suppliers, and real-time PCR was performed using the master mix from the QuantiTect Probe PCR +UNG Kit. ΔCT on the Y-axis indicates CT values for non-UNG-treated samples subtracted from CT values for UNG-treated samples. The UNG supplied with the QuantiTect Kit provided the greatest ΔCT (9-12 cycles). This indicates that QIAGEN UNG digests carryover PCR products more effectively than UNG from other suppliers.
Two-step RT-PCR.

The QuantiTect Probe PCR Kit overcomes the need for optimization of reaction conditions, which can be tedious and time-consuming. Simply add primers, probe, and cDNA template to the ready-to-use PCR master mix, and start the reaction on any real-time cycler (see also the table "Components of 2x QuantiTect Probe PCR Master Mix").
Highly specific amplification. In comparison with other DNA polymerases, only HotStarTaq DNA Polymerase in combination with a unique buffer specifically amplified a 497 bp fragment (from 50 copies of an HIV-pol-gene construct in a background of 1 µg human genomic DNA). M: Markers.
Wide dynamic range in two-step PCR.

Duplicate reactions were performed on the Mx3005P using tenfold dilutions of human keukocyte cDNA (10 ng to 0.01 ng) and a TaqMan assay for IL1R2 (a cytokine). The QuantiTect Probe PCR Kit provided accurate gene expression analysis from low to high template amounts with a PCR efficiency of 101%.
Wide dynamic range in real-time PCR. Probe-based real-time PCR with UNG pretreatment was carried out using the QuantiTect Probe PCR +UNG Kit. Reactions were run in duplicate on the ABI PRISM 7900 using 10-fold dilutions of human leukocyte cDNA (100 ng to 10 pg) and a TaqMan assay for IL8 (a cytokine). The amplification plots were evenly spaced, leading to a high PCR efficiency of 94%.
High sensitivity and efficiency, and wide dynamic range. Tenfold serial dilutions of leukocyte cDNA (100 ng to 1 pg) were analyzed in duplicate on the Mx3005 using the QuantiTect Probe PCR Kit with primers and a FAM-labeled probe specific for IL8 (interleukin 8). A PCR efficiency of 97% over 6 logs of template dilution was achieved, and as little as 1 pg of IL8 transcript was sensitively detected.