QuantiTect Multiplex RT-PCR Kits
For multiplex, one-step qRT-PCR using sequence-specific probes for gene expression analysis
- Multiplex analysis with no need for optimization
- Analysis of multiple targets in a single reaction
- Sensitive detection of as few as 10 copies of each target
- Reliable quantification of target and reference genes
- Detection of reference gene and up to 3 targets in the same tube
QuantiTect Multiplex RT-PCR Kits enable reliable quantification of up to 5 RNA targets in a single tube by multiplex, real-time one-step RT-PCR. The combination of a hot start and a unique PCR buffer system in the ready-to-use master mix ensures highly sensitive and reliable multiplex qRT-PCR on any real-time cycler without the need for optimization. The dNTP mix includes dUTP, allowing optional treatment with UNG. Two kit formats are available: the QuantiTect Multiplex RT-PCR Kit for cyclers that require ROX dye for fluorescence normalization, and the QuantiTect Multiplex RT-PCR NoROX Kit for all other cyclers. For convenience, the master mix can be stored at 2–8°C.
- Buy Products
- Product Details
- Specifications
Buy Products
Cat No./ID:204643 QuantiTect Multiplex RT-PCR Kit (200) $1,037.00 For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Multiplex RT-PCR Master Mix (with ROX dye), 100 µl QuantiTect Multiplex RT Mix, 2 x 2 ml RNase-Free Water |
Cat No./ID:204645 QuantiTect Multiplex RT-PCR Kit (1000) $4,079.00 For 1000 x 50 µl reactions: 25 ml 2x QuantiTect Multiplex RT-PCR Master Mix (with ROX dye), 0.5 ml QuantiTect Multiplex RT Mix, 20 ml RNase-Free Water |
Cat No./ID:204843 QuantiTect Multiplex RT-PCR NR Kit (200) $1,006.00 For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Multiplex RT-PCR NoROX Master Mix (without ROX dye), 100 µl QuantiTect Multiplex RT Mix, 2 x 2 ml RNase-Free Water |
Cat No./ID:204845 QuantiTect Multiplex RT-PCR NR Kit (1000) $4,026.00 For 1000 x 50 µl reactions: 25 ml 2x QuantiTect Multiplex RT-PCR NoROX Master Mix (without ROX dye), 0.5 ml QuantiTect Multiplex RT Mix, 20 ml RNase-Free Water |
Product Details
QuantiTect Multiplex RT-PCR Kits provide sensitive detection of as little as 10 copies of target sequence (see figure "Detection of down to 10 copies of target RNA in duplex PCR"). Optimal results are achieved even in 4-plex assays to quantify transcripts that strongly differ by up to 100,000-fold in starting copy number (see figure "Comparable amplification in 4-plex PCR and singleplex PCRs").
Precise relative quantification of the expression of a gene is achieved by quantifying the expression of both the target gene and an endogenous control gene in the same well or tube. With QuantiTect Multiplex RT-PCR Kits, the CT values for the targets in a multiplex reaction are equivalent to those obtained in control experiments where the targets are amplified in separate reactions (see figure "Comparable amplification in triplex PCR and singleplex PCRs"). This demonstrates that the different targets in the same multiplex reaction are efficiently and sensitively amplified without affecting each other.
Precise relative quantification of the expression of a gene is achieved by quantifying the expression of both the target gene and an endogenous control gene in the same well or tube. QuantiTect Multiplex RT-PCR Kits enable success in multiplex, one-step RT-PCR on the first attempt (see flowchart "QIAGEN multiplex kits"). The optimized master mix ensures that PCR products in a multiplex reaction are amplified with the same efficiency and sensitivity as PCR products in a corresponding single-amplification reactions. As few as 10 copies of a target gene can be detected with the kit.
Amplifying reference and target genes in the same reaction instead of in separate reactions increases the reliability of gene quantification by minimizing handling errors. The QuantiTect Multiplex RT-PCR Master Mix contains a balanced combination of K+ and NH4+ ions as well as unique synthetic Factor MP, which together promote stable and efficient annealing of primers and probes to the nucleic acid template, enabling high PCR efficiency (see figure "Unique PCRbuffer").In addition, anoptimized mix of reverse transcriptasesenables cDNA synthesis from a wide range of RNA template amounts, while HotStarTaq DNA Polymerase provides a stringent hot start, preventing the formation of nonspecific products.
The QuantiTect Multiplex RT-PCR Master Mix also contains dUTP, enabling pretreatment with uracil-N-glycosylase (UNG) prior to starting PCR, which ensures that any contaminating PCR products do not affect subsequent PCR reactions.
| Component | Features | Benefits |
|---|---|---|
| HotStarTaq DNA Polymerase | 15 minactivation at 95ºC | Set-up of qPCR reactions at room temperature |
| QuantiTect Multiplex RT-PCR Buffer | Balanced combination of NH4+ and K+ ions | Specific primer annealing ensures reliable PCR results |
| Synthetic Factor MP | Reliable multiplexing analysis of up to 4 genes in the same tube | |
| dNTP mix | Includes dUTP, which partially replaces dTTP and enables optional UNG treatment of reactions | Eliminatescontamination from carryover of PCR products by optional UNG treatment |
| ROX dye* | For normalization of fluorescent signals on Applied Biosystems and, optionally, Agilent instruments | Precise quantification on cyclers that require ROX dye.Does not interfere with reactions on other real-time cyclers |
| Omniscript and Sensiscript Reverse Transcriptases | Special blend of enzymes with high affinity for RNA | RNA can be transcribed, even through complex secondary structures |
QuantiTect Multiplex RT-PCR Kits contain ready-to-use master mixes that eliminate the need for optimization of reaction and cycling conditions. The handbook contains a single protocol that can be used with all available real-time cyclers and also lists recommended dyes. If required, reactions can be pretreated with uracil-N-glycosylase (UNG; not supplied) to eliminate carryover of PCR products from previous reactions.
Kits are available with or without ROX passive reference dye in the master mix, enabling use on virtually any real-time cycler (see table). Due to the optimized ROX concentrations, detection of even low copy numbers is achieved through automatic data analysis.
| ROX dye | Kit | Compatible cyclers |
|---|---|---|
| Supplied in master mix | QuantiTect Multiplex RT-PCR Kit | Cyclers from Applied Biosystems |
| Absent from master mix | QuantiTect Multiplex RT-PCR NR Kit | Rotor-Gene cyclers, and cyclers from Bio-Rad, Cepheid, Eppendorf, Roche, Agilent, and other suppliers |
QuantiTect Multiplex RT-PCR Kits can be used for gene expression analysis of RNA targets on any real-time cycler. This includes instruments from Applied Biosystems, Bio-Rad, Cepheid, Eppendorf, Roche, and Agilent. For the Rotor-Gene Q and other Rotor-Gene cyclers, we recommend using the Rotor-Gene Multiplex RT-PCR Kit, which has been specially developed for fast cycling on these instruments.
Specifications
Features | Specifications |
| Applications | Real-time quantification of RNA targets in a multiplex format |
| Description | For quantitative, multiplex, real-time one-step RT-PCR using sequence-specific probes |
| Reaction type | Real-time one-step RT-PCR |
| Real-time or endpoint | Real-time |
| Sample/target type | RNA targets |
| Single or multiplex | Multiplex |
| SYBR Green I or sequence-specific probes | Sequence-specific probes |
| Thermal cycler | Real-time cyclers dedicated for multiplex PCR (e.g., most Applied Biosystems real-time cyclers, Roche LightCycler 480, and Bio-Rad iCycler iQ) |
| With or without ROX | With ROX or without |
Product Resources
Customers who bought these products also bought
- Cat No./ID:940014
BioSprint 15 DNA Blood Kit (45)
For 45 preps: 5-Rod Covers, 5-Tube Strips, MagAttract Suspension G, Buffers and Reagents$170.00Add To Cart - Cat No./ID:32149
QIAexpress Type IV Kit
5 μg each: pQE-30, pQE-31, pQE-32 (N-terminal 6xHis); 10 ml Ni-NTA Agarose$741.00Add To Cart - Cat No./ID:11981
5 ml Tube Centrifuge Blocks
For easy centrifugation of MO BIO 5 ml bead tubes in standard 96 well centrifuge buckets.$205.00Add To Cart - Cat No./ID:204745
QuantiTect Multiplex PCR NoROX Kit (1000)
For 1000 x 50 µl reactions: 25 ml 2x QuantiTect Multiplex PCR NoROX Master Mix (without ROX dye), 20 ml RNase-Free Water$2,983.00Add To Cart - Cat No./ID:204543
QuantiTect Multiplex PCR Kit (200)
For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Multiplex PCR Master Mix (with ROX dye), 2 x 2 ml RNase-Free Water$732.00Add To Cart - Cat No./ID:290025
mericon L. monocytogenes Kit (96)
For 96 reactions: PCR Assay L. monocytogenes, Internal Control, Positive Control, Multiplex PCR Master Mix, QuantiTect Nucleic Acid Dilution Buffer, RNase-free water, 50x ROX Dye Solution$752.00Add To Cart - Cat No./ID:990399
Spin Column Adapter Rings (25)
Spin column adapter rings for use with spin columns that have screw caps; not required for all protocols$149.00Add To Cart - Cat No./ID:941557
BioSprint 96 DNA Plant Kit (576)
For 576 preps: Large 96-Rod Covers, 96-Well Microplates MP, S Blocks, MagAttract Suspension G, buffers and reagents$1,058.00Add To Cart - Cat No./ID:929510
QX RNA Alignment Marker (1.5 ml)
RNA alignment marker$76.80Add To Cart - Cat No./ID:204743
QuantiTect Multiplex PCR NoROX Kit (200)
For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Multiplex PCR NoROX Master Mix (without ROX dye), 2x2ml RNase-Free Water$754.00Add To Cart - Cat No./ID:202744
UCP HiFidelity PCR Kit (500)
For 500 x 25 µl PCR reactions: 5 x 1.3 ml UCP HiFidelity MasterMix (2x); 2 x 200 μl UCP Template Tracer, 25x; 1 x 50 μl UCP Master MixTracer, 125x; 4 x 1.9 ml UCP PCR Water
$643.00Add To Cart - Cat No./ID:204554
QuantiFast Probe RT-PCR +ROX Vial Kit (400)
For 400 x 25 µl reactions: 3 x 1.7 ml 2x QuantiFast Probe RT-PCR Master Mix (without ROX dye), 210 µl ROX Dye Solution, 100 µl QuantiFast RT Mix, 2 ml RNase-Free Water$716.00Add To Cart - Cat No./ID:239025
GelPilot 50 bp Ladder (100)
Ready-to-use DNA marker; 600 µl (100 lanes), 9 fragments: 50–500 bp$176.00Add To Cart - Cat No./ID:292145
mericon Horse Kit (96)
For 96 reactions: PCR Assay Horse, Internal Control, Positive Control, Multiplex PCR Master Mix, QuantiTect Nucleic Acid Dilution Buffer, RNase-Free Water, 50x ROX Dye Solution$1,065.00Add To Cart - Cat No./ID:981601
Rotor-Disc Heat Sealing Film (60)
60 films for sealing Rotor-Disc 100 or Rotor-Disc 72 discs$120.00Add To Cart - Cat No./ID:150243
REPLI-g FFPE Kit (25)
DNA Polymerase, Buffers, and Reagents for 25 x 50 μl whole genome amplification reactions$632.00Add To Cart - Cat No./ID:239085
GelPilot 1 kb Ladder (100)
Ready-to-use DNA marker; 600 µl (100 lanes), 9 fragments: 1000–10,000 bp$137.00Add To Cart - Cat No./ID:204343
QuantiTect Probe PCR Kit (200)
For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Probe PCR Master Mix, 2 x 2 ml RNase-Free Water$479.00Add To Cart - Cat No./ID:981301
Rotor-Disc 72 (24)
24 individually wrapped discs for 1728 reactions of 20–25 µl$264.00Add To Cart - Cat No./ID:204443
QuantiTect Probe RT-PCR Kit (200)
For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Probe RT-PCR Master Mix, 100 µl QuantiTect RT Mix, 2 x 2 ml RNase-Free Water$672.00Add To Cart
