The ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, ROX) is a ready-to-use reagent with all the essential components for quantitative real-time PCR (qPCR) except primers and templates. The master mix contains a highly stable hot-start Taq polymerase in an optimized buffer with dsDNA specific SYBR green fluorescent dye. Consequently, the ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, ROX) features high stability during storage, even at 37°C for weeks. In addition to high sensitivity and signal intensity, the ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, ROX) performs a low background/ high specificity qPCR results, as well as a better compatibility with fast PCR program. With inert smart blue contrast dye, the ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, ROX) is ready-to-use and greatly reduces pipetting errors, while largely improving the reproducibility of the process. The ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, ROX) also includes ROX reference dye for normalizing the fluorescent reporter signal in real-time quantitative PCR. This master mix allows sensitive, precise amplification, real-time tracking of the amplification process, and simultaneous quantification for targeted DNA molecules.
Detail
Description
The ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, ROX) is a ready-to-use reagent with all the essential components for quantitative real-time PCR (qPCR) except primers and templates. The master mix contains a highly stable hot-start Taq polymerase in an optimized buffer with dsDNA specific SYBR green fluorescent dye. Consequently, the ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, ROX) features high stability during storage, even at 37°C for weeks. In addition to high sensitivity and signal intensity, the ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, ROX) performs a low background/ high specificity qPCR results, as well as a better compatibility with fast PCR program. With inert smart blue contrast dye, the ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, ROX) is ready-to-use and greatly reduces pipetting errors, while largely improving the reproducibility of the process. The ExcelTaq™ 2X Fast Q-PCR Master Mix (SYBR, ROX) also includes ROX reference dye for normalizing the fluorescent reporter signal in real-time quantitative PCR. This master mix allows sensitive, precise amplification, real-time tracking of the amplification process, and simultaneous quantification for targeted DNA molecules.
Features
High Stability
Fast Hot Start
High Sensitivity
Low Background
Suitable for Fast Program
Smart Blue Contrast Dye
With ROX Reference Dye
Storage
Protect from light. Aliquot to avoid multiple freeze-thaw cycles. -20°C for 12 months
Other Products
HiDi® 2x PCR Master Mix
Product Info
Document
Product Info
HiDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and therefor, is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) that is also termed allele-specific amplification (ASA).HiDi® 2x PCR Master Mix – ready to use mix simplifies your PCR setup. Only target-specific primers and template need to be added as the mix contains all components for a successful and reliable PCR. This ensures reproducible results, significantly reduces set-up times and the risk of pipetting errors.This PCR mix is also available with a full-length Taq DNA polymerase with a nuclease domain, featuring 100% compatibility with hydrolysis probes (TaqMan® probes etc.).Benchmarking with products of competitors conducted by us and others show that the HiDi® DNA polymerase family is the first choice for highly selective PCRs, such as genotyping by allele-specific PCR, HLA genotyping, analysis of single CpG methylation sites or the detection of mutations in a high background of wild-type sequences. By using HiDi® DNA polymerase, less than 10 copies of a mutation can be detected in a background of >10.000 wild-type copies straight away without any other tedious assay optimization.It has also been shown that HiDi® DNA polymerase family is highly suitable for quality control in CRISPR-Cas or TALEN-based applications.Several independently conducted studies show that HiDi® DNA polymerase is ideally suited for use in asPCR in numerous research areas ranging from mutation detection to genome editing. (read more)For research use and further manufacturing.In case you are aiming to use our RUO products as components or for your development of e.g. an IVD medical device, please contact us.
Casestudies: HiDi® DNA Polymerase: Applications from mutation detection to genome editing (read more)
Example Primer Design
Matching vs. mismatching nucleotide is placed at the 3′-end of the primer for best discrimination results.
Example Results – There´s no accounting for taste
Cilantro: some people love it in their food, some hate it. Here we are detecting a genomic SNP (rs72921001) in HeLa genomic DNA. This SNP is reported to be close to a number of genes coding for olfactory receptors. (Reference: Eriksson N. et al. (2012), “A genetic variant near olfactory receptor genes influences cilantro preference.”)
Considering, that only the C-allele specific primer is extended and yielding in a specific amplicon, we can conclude a genetic predisposition in disliking cilantro, as this SNP is significantly associated with detecting a soapy taste to cilantro.
Allele-specific PCRs were performed from 1 ng/µl of HeLa gDNA in the presence of a realtime dye, indicating the amplification of the C-allele specific primer only. The A-allele specific primer is discriminated, thus not amplified up to 50 cycles.
PCR products were subsequently analysed on a 2.5% agarose gel. Specific product is visualized by ethidium bromide staining at the amplicon length of 109 bp.
Document
HiDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and therefor, is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) that is also termed allele-specific amplification (ASA).
HiDi® 2x PCR Master Mix – ready to use mix simplifies your PCR setup. Only target-specific primers and template need to be added as the mix contains all components for a successful and reliable PCR. This ensures reproducible results, significantly reduces set-up times and the risk of pipetting errors.
Buffer system mainly for stabilizing protein/enzyme and performance
B buffer
0.15ml
1 Tube
Mainly activated systems such as magnesium ions
Positive control template
0.1ml
1 Tube
Mainly the positive plasmid template is used to test the effectiveness of the kit
Positive control primer mix
0.06ml
1 Tube
Mainly the primer combination of the positive control template
Reagent Guide Manua
16T/bags,48T/Box
3 bags
Reagent technology of protein/enzyme system: freeze-dried powder, freeze-dried microspheres
Principle overview
This kit is based on a room temperature and constant temperature nucleic acid rapid amplification technology: at room temperature and constant temperature, the recombinase and primer form the protein/single-stranded nucleotide complex Rec/ssDNA, with the help of auxiliary proteins and single-stranded binding protein SSB , invade the double-stranded DNA template; form a D-loop region at the invasion site, and start scanning the DNA double-strands; after finding the target region complementary to the primer, the Rec/ssDNA complex disintegrates, and the polymerase also binds to The 3′ end of the primer initiates chain extension. This kit relies on the action of exonuclease at 39 ºC, adding specific molecular probes designed based on the template, and using fluorescence monitoring equipment to achieve real-time monitoring of the amplification process of the target fragment.
Primer design
It is recommended to use primers with a length of 30-35 bp. Primers that are too short will affect amplification speed and detection sensitivity; primers are designed to avoid the formation of secondary structures that affect amplification; the amplicon length is recommended to be 150-300 bp, usually no more than 500 bp.
Fluorescent probe design
The probe sequence does not overlap with the specific primer recognition site, is 46-52 nt in length, and the sequence avoids palindromic sequences, internal secondary structures, and continuous repeated bases. The probe has four modification sites: the middle position ≥ 35 nt from the 5′ end is labeled with a dSpacer (tetrahydrofuran, THF) as the recognition site for exonuclease; the upstream of the THF site is labeled with a fluorescent group, and the downstream Label a quenching group, the distance between the two groups is 2-4 nt; THF is ≥15 nt from the 3′ end, and the 3′ end is labeled with a modifying group, such as an amine group, a phosphate group or a C3-Spacer.
Product features and advantages:
This kit has the advantages of high sensitivity, strong specificity,and short reaction time (only 20 minutes), and the reaction groups are in dry powder state, which is easy to operate and easy to store.
It can be applied to various brands of fluorescence quantitative PCR instruments, constant temperature fluorescence amplification instruments and other fluorescence detection equipment.
Document
This kit is based on a room temperature and constant temperature nucleic acid rapid amplification technology: at room temperature and constant temperature, the recombinase and primer form the protein/single-stranded nucleotide complex Rec/ssDNA, with the help of auxiliary proteins and single-stranded binding protein SSB , invade the double-stranded DNA template; form a D-loop region at the invasion site, and start scanning the DNA double-strands; after finding the target region complementary to the primer, the Rec/ssDNA complex disintegrates, and the polymerase also binds to The 3′ end of the primer initiates chain extension. This kit relies on the action of exonuclease at 39 ºC, adding specific molecular probes designed based on the template, and using fluorescence monitoring equipment to achieve real-time monitoring of the amplification process of the target fragment.
CE-IVD marked version available for in vitro diagnostic use
Available in TaqMan format for analysis
Salmonella enterica have emerged as significant foodborne pathogens that pose a serious public health problem. The symptoms of salmonellosis may include diarrhea, fever, vomiting, and abdominal cramps with elderly, new-born, and immunocompromised individuals the most susceptible. S. enterica is a facultatively anaerobic Gram-negative bacterium that could survive low temperatures and freezing. The majority of the 1.3 billion annual cases of Salmonella-caused human gastroenteritis result from ingestion of contaminated food products, such as raw or undercooked meat, seafood, and eggs, as well as raw or unpasteurized milk and dairy products. Salmonella infections are also contracted following consumption of fresh fruits or vegetables that have been contaminated by infected fertilizer.
Salmonella enterica TaqMan PCR Kit, 100 reactions
Ready to use format, including Master Mix for the target and PCR control to monitor for PCR inhibition and validate the quality
Specific Primer and Probe mix for the pathogen/virus/viroid of interest
Primer and Probe mix
Positive and negative control to confirm the integrity of the kit reagents
Salmonella enterica TaqMan PCR Probe/Primer Set and Controls, 100 reactions
Specific Primer/Probe mix and Positive Control for the pathogen/virus/viroid of interest
Nuclease-free water
Can be used together with Norgen’s PCR Master Mix (#28007) or customer supplied master mix
Storage Conditions and Product Stability All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
