Hieff UNICON™ Universal Blue qPCR Master Mix (SYBR) _ 11184ES
Hieff UNICON™ Universal Blue qPCR Master Mix (SYBR) _ 11184ES
Hieff UNICON™ Universal Blue qPCR Master Mix (SYBR) _ 11184ES
Hieff UNICON™ Universal Blue qPCR Master Mix (SYBR) _ 11184ES
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Hieff UNICON™ Universal Blue qPCR Master Mix (SYBR) _ 11184ESHieff UNICON™ Universal Blue qPCR Master Mix (SYBR) _ 11184ESHieff UNICON™ Universal Blue qPCR Master Mix (SYBR) _ 11184ESHieff UNICON™ Universal Blue qPCR Master Mix (SYBR) _ 11184ES
Hieff UNICON™ Universal Blue qPCR Master Mix (SYBR) _ 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%),...

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Hieff UNICON™ Universal Blue qPCR Master Mix (SYBR) _ 11184ES

YeasenSKU: 11184ES03
Size: 1 mL
Price:
$55.00
Stock:
In stock

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.

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

Applications

Gene Expression

Specifications

Concentration 2×
Detection method SYBR Green dye
PCR method qPCR
Polymerase Taq DNA polymerase
Type of sample DNA
Application equipment Most qPCR instruments
Product Type Premix for real-time fluorescence quantitative PCR
Apply to (application) Gene Expression

Components

Components No. Name 11184ES03 11184ES08 11184ES50 11184ES60
11184

Hieff Unicon™ Universal Blue qPCR Master Mix (Dye Based)

 1 mL 5×1 mL 50×1 mL 100×1 mL

Figures

Figure 1. High sensitivity: the ability to detect a single copy.


Figure2. High resolution & Excellent reproducibility of duplicate wells.

Figure3. Excellent detection rate and specificity: broadly suited for the amplification of 30-70% GC.

Cited from "VCP enhances autophagy-related osteosarcoma progression by recruiting USP2 to inhibit ubiquitination and degradation of FASN, Cell Death & Disease, volume 15, Article number: 788 (2024) "

Cited from "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. "

Storage

The product is shipped with ice packs and can be stored at -15℃ ~ -25℃ for 18 months. The product contains fluorescent dyes, so it is necessary to avoid strong light irradiation when storing or preparing the reaction system.

FAQ

 The common method involves two steps. To enhance amplification specificity, one can choose the two-step method or increase the annealing temperature. When the amplification efficiency is low and the ct value is too high, a three-step method can be adopted or the extension time can be prolonged.
The validity must meet the following three conditions:
(1) Standard curve: Amplification efficiency range: 90-110%, corresponding slope: -3 to -3.5. R2 > 0.98. (Amplification efficiency = 10 - 1/slope - 1), when the slope is -3.32, the amplification efficiency is 100%.
(2) Amplification curve: S-shaped curve, and Ct value is between 15 and 35, negative control Ct > 35 or no Ct value.
(3) Melting curve: Single peak. The Ct value is greater than 15 cycles because the 10 times standard deviation of the fluorescence value in 3-15 cycles is the fluorescence threshold, and a too small Ct value will affect the curve.
Single copy
Even for the same amplified product, there will be slight differences in the Tm values. Generally, a difference of less than 1 degree is acceptable.
Generally, the CT value is negatively correlated with the initial concentration of the template. The higher the concentration, the lower the CT value. However, there are also many special cases. For instance, if there are inhibitors in the system or the template is not pure, diluting the template can actually result in a lower CT value.
This might be due to the low expression level of the target gene. Suggestions:
a) Use a different reference gene;
b) Change the primers.
The possible reason is that the ROX was added improperly. Verify whether the reference dye ROX was added correctly.
(1) The amount of template DNA needs to be determined by the experimenter during the initial experiment. Firstly, dilute the template DNA (generally recommended to dilute by 5-10 times), then sample at different template quantities on a gradient, and select the optimal sample amount with a CT value ranging from 20 to 30.
(2) The commonly used amount is 500-1000 ng of total RNA for reverse transcription, dilute it 10 times and take 1 μL of cDNA for the qPCR experiment.
a) The validity must meet the following three conditions:
(1) Standard curve: Amplification efficiency range: 90-110%, corresponding slope: -3 to -3.5. R2 > 0.98. (Amplification efficiency = 10 - 1/slope - 1), when the slope is -3.32, the amplification efficiency is 100%.
(2) Amplification curve: S-shaped curve, and Ct value is between 15 and 35, negative control Ct > 35 or no Ct value.
(3) Melting curve: Single peak.
b) The 10 times the standard deviation of the fluorescence values for 3 to 15 cycles is the fluorescence threshold. A too small Ct value will affect the curve.
ROX is a reference dye. Its function is to standardize the non-PCR oscillations in the fluorescence quantitative reaction, correct the sample addition errors or the errors between wells, and provide a stable baseline.
The reagent contains stabilizing components that do not participate in any reactions themselves, but they will show a diffused pattern on the electrophoresis gel. It is not recommended to perform electrophoresis after the run is complete.
You can first mix the large sample, then add it separately to the centrifuge tubes, and finally add the template. When adding samples to the gun head, make sure not to let any air in. Use the second gear for sample aspiration and the first gear for sample injection; slowly push out the liquid and do not blow forcefully; guide the liquid into the hole along the wall. After the configuration is completed, if there are still bubbles, remove the centrifuged PCR tube. If it is a 96-well plate, gently tap the 96-well plate after the pipetting is done.

Citations

Documents

Brochure
Comprehensive_And_Professional_qPCR_Solution.pdf
Hieff_qPCR_Master_Mix_Series.pdf

Applications

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Inquiry

FAQ

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