qPCR Reagents
Hieff Unicon™ Universal Blue qPCR Master Mix (SYBR Green Based) - 11184ES
Hieff Unicon™ Universal Blue qPCR Master Mix (Dye Based) is a pre-solution for 2×real-time quantitative PCR amplification characterized by high sensitivity and specificity, is blue in color, and has the effect of sample addition tracing. The core component Hieff Unicon™ Taq DNA polymerase uses antibody hot start to effectively inhibit non-specific amplification due to primer annealing during sample preparation. At the same time, the formula adds the promoting factors to improve the amplification efficiency of PCR reaction and equalize the amplification of genes with different GC contents (30 ~ 70%), so that quantitative PCR can obtain a good linear relationship in a wide quantitative region. This product contains special ROX Passive Reference Dye, which is applicable to most qPCR instruments. It is not necessary to adjust the concentration of ROX on different instruments. It is only necessary to add primers and templates to prepare the reaction system for amplification.
Figure 1. High sensitivity: the ability to detect a single copy.
Figure 2. High resolution & Excellent reproducibility of duplicate wells.
Figure 3. Excellent detection rate and specificity: broadly suited for the amplification of 30-70% GC.
Selected citations:
[1] SARS-CoV-2 envelope protein causes acute respiratory distress syndrome (ARDS)-like pathological damages and constitutes an antiviral target. Cell Res. 2021;31(8):847-860. doi:10.1038/s41422-021-00519-4
[2] A Defense Pathway Linking Plasma Membrane and Chloroplasts and Co-opted by Pathogens. Cell . 2020 Sep 3;182(5):1109-1124.e25. doi: 10.1016/j.cell.2020.07.020.
Features:
- Built-in visual indicator: It contains an inert blue dye that does not interfere with real-time PCR and provides higher visibility in tubes or Wells
- High specificity: It contains Taq DNA polymerase and is tightly controlled by an antibody-mediated heat-priming mechanism, thereby avoiding nonspecific amplification
- Wide instrument compatibility: It is compatible with most qPCR instruments and eliminates the need to adjust the concentration of ROX on different instruments
Hieff Unicon™ Universal Multiplex qPCR Master Mix (Probe Based) - 11211ES
A pre-solution for 2×real-time quantitative PCR amplification characterized by high sensitivity and specificity, is blue in color, and has the effect of sample addition
This product is a 2× Mix pre-mixed reagent that enables up to four fluorescent quantitative PCR reactions in a single reaction well. This product contains the genetically modified antibody method to hot-start Taq enzyme, greatly improving the amplification sensitivity and specificity. At the same time, this product has deeply optimized the multi-reaction buffer, which can improve the amplification efficiency of the reaction and promote the effective amplification of low-concentration templates. This product can be used for genotyping and multiplex quantitative analysis. Figure 1. High stability: 37℃ thermal acceleration stability. The red line: 37℃ thermal acceleration 0 day; The yellow line: 37℃ thermal acceleration 7 days; The blue line: 37℃ thermal acceleration 14 days Figure 2 High stability: freeze-thaw stability. The red line: freeze thaw o time; The yellow line: freeze thaw 50 times; The blue line: NTC Selected citations: [1] Transcriptomic and Metabolomic Profiling Reveals the Effect of LED Light Quality on Fruit Ripening and Anthocyanin Accumulation in Cabernet Sauvignon Grape. Front Nutr. 2021;8:790697 [2] Histone methyltransferase Dot1L recruits O-GlcNAc transferase to target chromatin sites to regulate histone O-GlcNAcylation. J Biol Chem. 2022;298(7):102115. doi:10.1016/j.jbc.2022.102115(IF:5.157)
This product is a 2× Mix pre-mixed reagent that enables up to four fluorescent quantitative PCR reactions in a single reaction well. This product contains the genetically modified antibody method to hot-start Taq enzyme, greatly improving the amplification sensitivity and specificity. At the same time, this product has deeply optimized the multi-reaction buffer, which can improve the amplification efficiency of the reaction and promote the effective amplification of low-concentration templates. This product can be used for genotyping and multiplex quantitative analysis. Figure 1. High stability: 37℃ thermal acceleration stability. The red line: 37℃ thermal acceleration 0 day; The yellow line: 37℃ thermal acceleration 7 days; The blue line: 37℃ thermal acceleration 14 days Figure 2 High stability: freeze-thaw stability. The red line: freeze thaw o time; The yellow line: freeze thaw 50 times; The blue line: NTC Selected citations: [1] Transcriptomic and Metabolomic Profiling Reveals the Effect of LED Light Quality on Fruit Ripening and Anthocyanin Accumulation in Cabernet Sauvignon Grape. Front Nutr. 2021;8:790697 [2] Histone methyltransferase Dot1L recruits O-GlcNAc transferase to target chromatin sites to regulate histone O-GlcNAcylation. J Biol Chem. 2022;298(7):102115. doi:10.1016/j.jbc.2022.102115(IF:5.157)
Features: Applications:
- Strong tolerance of blood impurities
- High sensitivity
- Superior storage stability
- Pathogen detection
- Copy number analysis
- SNP genotyping
Hifair™ V Multiplex One Step RT-qPCR Probe Kit （UDG Plus) - 13650ES
A multiplex quantitative PCR kit based on RNA as template. In the process of the experiment, reverse transcription and quantitative PCR were carried out in the same tube, which simplified the experimental operation and reduced the risk of contamination.
In this kit, the first strand cDNA was efficiently synthesized by heat-resistant Hifair™ V Reverse Transcriptase and quantitatively amplified by UNICON™ HotStart Taq DNA Polymerase. The kit mainly contains optimized MP buffer, enzymes mix, etc. The buffer solution already contains Mg2+ and dNTP. In addition, the factors that can effectively inhibit the non-specific PCR amplification and improve the amplification efficiency of multiple qPCR reactions are added, which can ensure the amplification efficiency and carry out up to multiple amplification reaction. The dUTP/UDG system was added to effectively prevent the risk of aerosol contamination.
In this kit, the first strand cDNA was efficiently synthesized by heat-resistant Hifair™ V Reverse Transcriptase and quantitatively amplified by UNICON™ HotStart Taq DNA Polymerase. The kit mainly contains optimized MP buffer, enzymes mix, etc. The buffer solution already contains Mg2+ and dNTP. In addition, the factors that can effectively inhibit the non-specific PCR amplification and improve the amplification efficiency of multiple qPCR reactions are added, which can ensure the amplification efficiency and carry out up to multiple amplification reaction. The dUTP/UDG system was added to effectively prevent the risk of aerosol contamination.
Validated equipment:
- Equipment with Rox：ABI 5700, 7000, 7300, 7700, 7900HT Fast, StepOne™, StepOne Plus™
- Equipment with Low Rox：ABI 7500, 7500 Fast, ViiA™7, QuantStudio™ 3 and 5, QuantStudio™ 6,7,12k Flex
- Stratagene MX3000P™, MX3005P™, MX4000P™
- Equipment without Rox: Bio-Rad CFX96™, CFX384™, iCycler iQ™, iQ™5, MyiQ™, MiniOpticon™, Opticon®, Opticon® 2, Chromo4™
- Equipment without Rox: Eppendorf Mastercycler® ep realplex, realplex 2 s; Qiagen Corbett Rotor-Gene® Q, Rotor-Gene® 3000, Rotor-Gene® 6000
- Equipment without Rox: Roche Applied Science LightCycler® 480, LightCycler® 2.0, Lightcycler® 96
- Equipment without Rox: Thermo Scientific PikoReal Cycler; Cepheid SmartCycler®; Illumina Eco qPCR
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Hifair® Advanced One Step RT-qPCR SYBR Green Kit - 11175ES
A kit for fluorescence quantification based on SYBR Green I dye. Using gene-specific primers, reverse transcription and qPCR reactions are completed in one tube, eliminating the need for repeated cap-opening and pipetting operations, greatly improving assay efficiency and reducing the risk of contamination. For RNA samples, the kit employs heat-resistant Hifair™ V Reverse Transcriptase for efficient cDNA synthesis and UNICON™ HotStart Taq DNA Polymerase for quantitative amplification. Under the optimized buffer system, the sensitivity of the kit can be as high as 0.1 pg for highly expressed targets and as high as 1 pg for moderately expressed targets, and the kit is also suitable for amplification and quantification of DNA samples. The kit is suitable for the amplification and quantification of DNA samples. It enables the sensitive detection and quantification of nucleic acids from different plant and animal samples, cells and microorganisms.
Validated equipment:
- Instrument models that do not require Rox calibration: Bio-Rad: CFX96, CFX384, iCycler iQ, iQ5, MyiQ, MiniOpticon, Opticon, Opticon 2, Chromo4; Eppendorf: Mastercycler ep realplex, realplex 2 s; Qiagen: Corbett Rotor-Gene Q, Rotor-Gene 3000, Rotor-Gene 6000; Roche Applied Science: LightCycler 480, LightCycler2.0, Lightcycler 96; Thermo Scientific: PikoReal Cycler; Cepheid: SmartCycler; Illumina: Eco qPCR;
- Low Rox applicable models: ABI 7500, 7500 Fast, ViiA7, QuantStudio 3 and 5, QuantStudio 6, 7, 12k Flex; Stratagene MX3000P, MX3005P, MX4000P;
- High Rox applicable models: ABI 5700, 7000, 7300, 7700, 7900HT Fast, StepOne, StepOne Plus.
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Hieff® Fast Cell Direct SYBR Green RT-qPCR Kit - 11172ES
Hieff™ Fast Cell Direct SYBR Green RT-qPCR Kit is suitable for RNA extraction from all kinds of animal cells (such as cell lineage wall cells and suspension cells, primary culture cells, various stem cells, iPS cells, etc.), without the need to extract RNA, and can be used directly for qPCR expression analysis, which is short, easy to operate, and has a low error rate. It takes only 1.5 hours to complete the steps from template preparation to reverse transcription reaction and gene expression analysis.
Reverse transcription and fluorescence detection reagents are included in the kit, which can be used for gene expression analysis without the need to purchase additional reagents.
Reverse transcription and fluorescence detection reagents are included in the kit, which can be used for gene expression analysis without the need to purchase additional reagents.
Features:
- Time saving: Saving 3 hours
- Direct RT-qPCR: Facilitating low copy gene detection
- Convenient: Without RNA extraction
- Fast: Completed in 50 minutes
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Hieff™ miRNA Universal qPCR SYBR Master Mix (by poly A) - 11171ES
MicroRNA, a class of non-coding RNAs with a length of about 22 nt, is widely concerned and plays a very important role in regulating gene expression in plants and animals. This kit uses the SYBR Green I chimeric fluorescence method for the quantitative detection of miRNA fluorescence. The 2×Hieff™ miRNA Universal qPCR SYBR Master Mix is a new generation of premix fluorescent quantitative PCR detection reagent specially developed for the quantitative detection of miRNA. It contains special ROX Passive Reference Dye. Suitable for most qPCR instruments, there is no need to adjust the concentration of ROX on different instruments, just add primers and templates in the preparation of the reaction system can be amplified. The DNA Polymerase uses chemically modified heat-activated Polymerase, combined with a special Buffer system, to make the reaction more specific, more sensitive, and can be accurately quantified in a wider range.This product is recommended for use with our Hifair™ miRNA 1st Strand cDNA Synthesis Kit (Tailing) (Cat# 11148) to obtain optimal experimental results.
Figure1. Good amplification performance: excellent amplification efficiency. miR-let-7b-5p, miR-let-7c-5p, miR-let-7e-5p, and miR-let-7f-5p genes were amplified using human 293T cells and mouse liver Total RNA as templates. The results showed that compared with similar products, Hifair™ miRNA 1st Strand cDNA Synthesis Kit (Cat# 11148ES) matched the reaction system of Hieff™ miRNA Universal qPCR SYBR Master Mix (Cat# 11171ES) with excellent performance.
Figure2. High specificity: be able to distinguish single base differences between miRNAs in the same family. The human hsa-let-7 family has multiple miRNAs, with few bases that differ from each other, or even only a single base difference. Amplify these miRNAs with different primers and use Formula 2- Δ CT x 100% calculated matching rate. The results showed that closely related subtype family members could be effectively distinguished.
Features:
- Platform-wide: No need to alter ROX concentration using premixed colors
- High sensitivity detection rate: 10 pg DNA can be detected
- Good amplification performance: Excellent amplification efficiency
- High specificity: Able to distinguish single base differences between miRNAs in the same family
Hieff Unicon™ qPCR TaqMan Probe Master Mix - 11205ES
The product is a premixed solution for 2× real-time quantitative PCR amplification by the probe method. The antibody hot-start UNICONTM DNA Polymerase used can effectively inhibit the amplification caused by the non-specific annealing of primers. At the same time, the formula optimizes the factors that can effectively inhibit non-specific PCR amplification and improve the amplification efficiency of PCR reaction, which is suitable for the amplification of low-concentration templates, so that quantitative PCR can obtain a good standard curve in a wide quantitative area.
Selected citations:
[1] The CREB/KMT5A complex regulates PTP1B to modulate high glucose-induced endothelial inflammatory factor levels in diabetic nephropathy. Cell Death Dis. 2021;12(4):333. Published 2021 Mar 29. doi:10.1038/s41419-021-03629-4(IF:8.469)
[2] Formaldehyde reinforces pro-inflammatory responses of macrophages through induction of glycolysis. Chemosphere. 2021;282:131149. doi:10.1016/j.chemosphere.2021.131149(IF:7.086)
Validated equipment:
- Applied Biosystems: 5700, 7000, 7300, 7700, 7900HT Fast, StepOne™, StepOne™ Plus; 7500, 7500 Fast, ViiA™7, QuantStudio™ 3 and 5, QuantStudio™ 6, 7, 12k Flex; Bio-Rad：CFX96, CFX384, iCycler iQ, iQ5, MyiQ, MiniOpticon, Opticon, Opticon 2, Chromo4; Eppendorf：Mastercycler ep realplex, realplex 2 s; Qiagen：Corbett Rotor-Gene Q, Rotor-Gene 3000, Rotor-Gene 6000; Roche Applied Science：LightCycler 480, LightCycler 2.0; Lightcycle r 96; Thermo Scientific：PikoReal Cycler; Cepheid：SmartCycler; Illumina: Eco qPCR.
