RNA into DNA and PCR in one step? Then, this enzyme will simplify PCR analysis from RNA templates reducing labor and time. RT-KTQ2 was evolved from the thermostable KlenTaq DNA polymerase with no significant reverse transcriptase activity. Four mutations ensure that the variant is reverse transcriptase active and even PCR active, while maintaining the thermostability. This allows to perform reactions at high temperatures minimizing problems encountered with strong secondary structures in RNA that melt at elevated temperatures. For further information refer to the original publication.
Detail
RNA into DNA and PCR in one step? Then, this enzyme will simplify PCR analysis from RNA templates reducing labor and time. RT-KTQ2 was evolved from the thermostable KlenTaq DNA polymerase with no significant reverse transcriptase activity. Four mutations ensure that the variant is reverse transcriptase active and even PCR active, while maintaining the thermostability. This allows to perform reactions at high temperatures minimizing problems encountered with strong secondary structures in RNA that melt at elevated temperatures. For further information refer to the original publication.
Available upon request and for R&D use only – Contact Us
RT-KTQ2 DNA polymerase is supplied as a 5 µM solution containing glycerol and is supplied together with 10x reaction buffer.
The enzyme can also be used for real-time cycling, when adding a suitable dye.
The IdPath™ COVID-19 Real-Time RT-PCR Kit is a real-time RT-PCR test intended for the detection of SARS-CoV-2 virus RNA which might be extracted from the respiratory tract specimens. The Kit provides reagents for multiplex real-time RT-PCR to detect SARS-CoV-2 by one-step reaction, specifically targeting the E (Envelope), RdRP (RNA-dependent RNA polymerase) and N (Nucleocapsid protein) gene for SARS-CoV-2 virus.
The Kit contains the RT enzyme mix and qPCR master mix for reverse transcription and real-time PCR of virus RNA. The COVID-19 Control (positive control) and ddWater (negative control) are used as indicators to avoid false negative/positive results across all experimental procedures. The Primers/probes Mix contains multiplex primers and TaqMan probes specific to the N, E/RdRP genes of SARS-CoV-2 and RNase P gene of human, detected by FAM, VIC and ROX channels, respectively.
For Research Use Only
Features
High Sensitivity：5×102 copies/ml (10 copies/rxn)
High Inclusivity : >99% of currently available complete virus genomes for SARS-CoV-2 including Omicron variant
High Accuracy : Clinical validation with 100% accuracy
High Stability : 37/25°C for 2 weeks ; 4°C for 24 weeks ; 10 times of freeze-thaw cycles
High Compatibility : Suitable for most laboratory qPCR machines
Operation Control : Including internal control for quality control of total process
Convenience : Multiplex (E/RdRP and N) detection by one-step reaction
Storage
Aliquot to avoid multiple freeze-thaw cycles
Protect fluorogenic probes from light
-20°C for 12 months
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The IdPath™ COVID-19 Real-Time RT-PCR Kit is a real-time RT-PCR test intended for the detection of SARS-CoV-2 virus RNA which might be extracted from the respiratory tract specimens. The Kit provides reagents for multiplex real-time RT-PCR to detect SARS-CoV-2 by one-step reaction, specifically targeting the E (Envelope), RdRP (RNA-dependent RNA polymerase) and N (Nucleocapsid protein) gene for SARS-CoV-2 virus. The Kit contains the RT enzyme mix and qPCR master mix for reverse transcription and real-time PCR of virus RNA. The COVID-19 Control (positive control) and ddWater (negative control) are used as indicators to avoid false negative/positive results across all experimental procedures. The Primers/probes Mix contains multiplex primers and TaqMan probes specific to the N, E/RdRP genes of SARS-CoV-2 and RNase P gene of human, detected by FAM, VIC and ROX channels, respectively.
Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.
The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected*. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit.
Details of the target and priming specificity are included in the individual handbooks above.
Packaged, optimised and ready to use. Expect Better Data.
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Exceptional value for money Rapid detection of all clinically relevant subtypes Positive copy number standard curve for quantification Highly specific detection profile High priming efficiency Broad dynamic detection range (>6 logs) Sensitive to < 100 copies of target
Accurate controls to confirm findings
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 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).This polymerase is also available as 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.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.
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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 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).
