Products >> Lifescience >> Molecular Biology Reagents >> Kapa Probe Fast qPCR Kits
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KAPA Probe Fast qPCR Kits

KAPA PROBE FAST qPCR Kits contain a ready-to-use master mix for highly sensitive and accurate real-time PCR using sequence-specific probe chemistries including TaqMan®, FRET probes, and molecular beacons.

Optimized for versatility and speed – KAPA PROBE FAST qPCR Kits provide fast and reproducible results for genotyping, gene expression analysis, and multiplexing offering:

	Compatibility with all probe-based qPCR applications and instruments
	Fast, reproducible, and precise quantification
	Discrete clusters in SNP genotyping assays
	Multiplex qPCR
	Broad dynamic range
	Highly stable master mix for high-throughput workflows

Precise, reproducible, and versatile for all probe-based qPCR applications.

Discrete clusters and high call rates for accurate and reproducible allelic discrimination.

All 168 human genomic DNA samples were accurately genotyped using the ABI sequence detection system (SDS) version 2.3 software (autocaller confidence level 95%). A total of 168 human genomic DNA samples were successfully genotyped along with 24 no-template controls using an ATP1B3 SNP genotyping assay on the ABI 7900HT real-time PCR system. Reactions were performed in 5 μl volumes with KAPA PROBE FAST qPCR Master Mix, human genomic DNA (10 ng per reaction), 200 nM of each primer and 200 nM of each hydrolysis probe (Allele X – FAM/ BHQ®-1, Allele Y – VIC/ BHQ®-1).

Fast, high performance five-color multiplexed qPCR.

Highly reproducible and efficient results for all 5 amplicons across a 5 point dilution series of human genomic DNA were obtained when assayed in penta-plex using a fast cycling protocol. Standard curves were generated using 4-fold dilutions of human genomic DNA (0.39 – 100 ng per reaction) tested in triplicate using the Corbett Rotor-Gene™ 6000 HRM real-time rotary analyzer. Reactions were performed in 20 μl volumes with KAPA PROBE FAST qPCR Master Mix, human genomic DNA, 200 nM of each primer and 200 nM of each hydrolysis probe (ACTB - FAM™/BHQ®-1, ERBB2 - CAL Fluor® Gold 540/ BHQ®-1, ERBB3 - CAL Fluor® Orange 560/ BHQ®-2, EGFR - CAL Fluor® Red 610/ BHQ®-2, ACTG1 - Quasar® 705/ BHQ®-2).

High performance with both standard and fast cycling protocols.

Excellent reproducibility and efficiency was achieved using both cycling protocols. Reactions were performed in 20 μl volumes with KAPA PROBE FAST qPCR Master Mix, human genomic DNA (10-fold dilutions over a 0.1 - 100 ng per reaction range), 200 nM of each primer and 200 nM of hApoB100 (FAM/ BHQ®-1) hydrolysis probe using followed by 40 cycles of 95ºC, 15 sec; 60 ºC, 60 sec) or a fast cycling protocol (95 ºC for 3 min followed by 40 cycles of 95 ºC, 3 sec; 60 ºC, 20 sec).

Precise and highly reproducible low copy number discrimination.

Reproducibility and discrimination across samples of low copy number with similar abundance levels was achieved. Reactions were performed in 20 μl volumes with KAPA PROBE FAST qPCR Master Mix, human genomic DNA (1.5-fold dilutions over a 89 - 450 copies per reaction range), 200 nM of each primer and 200 nM of hApoB100 (FAM/ BHQ®-1) hydrolysis probe using a fast cycling protocol (95 ºC for 3 min followed by 40 cycles of 95 ºC 3 sec; 60 ºC, 20 sec).

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