4X 一步法 RT-qPCR 预混型探针法试剂盒（含 UNG）

4X One-Step RT-qPCR Pre-Mixed Probe Assay Kit (with UNG)

Product Code: CM0270

- +

¥1,800

Product Description

This is a specialized kit for one-step RT-qPCR based on the probe method. Reverse transcription and qPCR reactions are completed in the same tube, which makes the operation simple and fast.
This product utilizes a new type of heat-resistant, high extension capacity Evo Super M-MLV Reverse transcriptase, integrated mutant Accurate Super Taq The superior performance of HS polymerase enables efficient cDNA synthesis and stable qPCR amplification in a short period of time. The product is widely used, with high impurity tolerance, and can amplify genes of different complexity, while it can be used for single or multiplexed detection. It is suitable for the detection of DNA/RNA and other trace viruses.
The dUTP/UNG anti-contamination system is introduced in this product. dUTP replaces dTTP in the PCR reaction, and the UNG enzyme selectively hydrolyzes dU-containing DNA strands without affecting non-dU-containing DNA strands, removing dU-containing contaminating templates introduced during the preparation of the PCR reaction system, thus effectively preventing the generation of false-positive results in PCR; At the same time, monoclonal antibody that can inhibit the activity of DNA Polymerase at room temperature is added to this product, which can effectively inhibit non-specific amplification, improve the sensitivity of the reaction and enhance the accuracy of the results.

Product Advantages

1. This is a one-step RT-qPCR reaction kit, which can complete the reverse transcription and qPCR reaction in a single tube, simplifying the operation, improving the efficiency, and effectively reducing the possibility of contamination caused by multiple operations.
2. This product adopts a new type of heat-resistant (up to 55℃), strong elongation ability. Evo Super M-MLV Reverse transcriptase and mutant Accurate Super Taq HS, paired with a carefully optimized reaction system, is characterized by high specificity and amplification efficiency.
3. The product is highly sensitive and can stably detect single-digit copy templates, and can perform single or multiple detection.
4. This product can perform DNA/RNA co-detection.
5. dUTP/UNG is a contamination prevention system that removes contamination of dU-containing PCR products, thus preventing false-positive PCR results and improving the accuracy of the results.

Product Composition
individual parts making up a compound norm
4X One Step Probe Mixture ( UNG Plus ) 1.25 ml

*: contains Evo Super M-MLV RTase Enzyme,Accurate Super Taq HS DNA Polymerase, UNG Enzyme and RNase Inhibitor, etc.

Preservation and transportation
Storage temperature: -20℃ storage
Transportation temperature:Dry ice transportation or -20°C ice pack transportation.
Example 1

Total RNA (100 ng-10 pg) from 293T cells was used as a template, and RNase free water was used as a negative control instead of template for triple detection of TFR gene (GC content of 38%, FAM channel), Actin gene (GC content of 59%, HEX channel), RPP30 gene ( GC content was 62%, CY5 channel). The quantification instrument used was ABI QuantStudio.TM 5 Real-Time PCR Systems. the results of the experiments were as follows:

The results are shown above:
1, different concentrations of templates can get good amplification within 45 cycles;
2. Negative controls were not detected within 45 cycles;
3, the amplification efficiency was 97.41 TP3T, R2= 0.9965.

The results are shown above:
1, different concentrations of templates can get good amplification within 45 cycles;
2. Negative controls were not detected within 45 cycles;
3, the amplification efficiency was 93.41 TP3T, R2= 0.9998.

 

The results are shown above:
1, different concentrations of templates can get good amplification within 45 cycles;
2. Negative controls were not detected within 45 cycles;
3, the amplification efficiency was 100.1%, R2= 0.9980.

 

Example 2

The human nasal C RNA virus, human glandular 7 DNA virus and human parainfluenza 1 RNA virus standards were used as templates for triple detection. 8 copies of human nasal C RNA virus template, 20 copies of human glandular 7 DNA virus template and 1 copy of human parainfluenza 1 RNA virus template were added to 20 μl of the reaction system, and 8 replicate wells were tested for each of the three types of viruses. All three viruses were detected in 8 replicate wells, and RNase free water was used as a negative control instead of template. The quantification instrument used was ABI QuantStudio.TM 5 Real-Time PCR Systems. the results of the experiments were as follows:

The results are shown above:
1, this product for low copy of the virus template, can be well detected, high sensitivity and good reproducibility.
2, This product can detect DNA / RNA virus template at the same time.
3. Negative controls were not detected within 45 cycles.

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