Qihengxing Blood Direct qPCR Kit is specially designed for blood samples. It does not require cumbersome nucleic acid extraction steps, and only requires simple lysis for qPCR amplification. It is suitable for a variety of sample types such as fresh blood, 4℃ stored blood, -20℃ frozen blood, and EDTA anticoagulated blood. It is compatible with Taqman probe method and SNP detection, easy to operate, accurate results, and the whole process from blood to genotyping/quantitative analysis can be comp
With the rapid development of molecular diagnostic technology, based on the requirements of clinical testing and scientific research applications for rapid, efficient and low-cost, direct blood amplification (blood direct amplification) technology without nucleic acid extraction has gradually become an important development trend. However, in existing blood direct amplification products, there are generally core problems such as insufficient lysis efficiency leading to reduced amplification sensitivity, narrow sample application range (cannot cover a variety of samples such as fresh blood, refrigerated blood and frozen blood), and poor compatibility with primer/probe systems. The key point of these problems is how to ensure the reliability and consistency of test results while simplifying the operating process.
Based on its advantages in nucleic acid amplification technology and reagent development, Qihengxing Bio has developed the StarLighter blood direct amplification probe method qPCR kit. By optimizing the lysis system and amplification conditions, the kit achieves efficient processing of various types of blood samples (compatible with complex sample models such as EDTA anticoagulated blood and frozen blood, and at the same time, it shows excellent adaptability to primers/probes with large differences in GC content and wide Tm value spans), and can directly perform qPCR detection without complex nucleic acid extraction steps. Its test results are comparable to traditional nucleic acid extraction methods in terms of fluorescence signal intensity and Ct value, while significantly improving the convenience of operation and detection speed, providing an innovative solution for blood direct amplification technology that combines accuracy and efficiency.
Product Introduction
This kit can be applied to various types of blood samples. It does not require tedious nucleic acid extraction and purification. The qPCR amplification can be performed by simply lysing the blood sample and taking the supernatant. It is easy to operate and can reduce the risk of contamination. The test results are consistent with those using the pure DNA product after nucleic acid extraction as a template.
Product Advantages
1. Easy operation: heat at 95℃ for 5 minutes and then centrifuge. No complicated blood nucleic acid extraction steps are required.
2. High lysis and release efficiency: The lysis effect of the release agent is consistent with the effect of nucleic acid genome extraction from blood;
3. Wide sample compatibility: suitable for fresh blood, 4℃ stored blood, -20℃ frozen blood, EDTA anticoagulated blood and other samples;
4. Wide compatibility of primers/probes: ideal amplification can be achieved for primers/probes with different GC%/Tm values.
Application
1. Taqman probe method SNP detection;
2. Quantitative detection by qPCR probe method.
Product Performance
- Simple and fast operation.
It only takes about 1 hour from blood to genotyping/quantification analysis.
Fig.1 Kit operation flow chart
- Accurate SNP typing, compatible with fresh blood, 4 ℃ Storage blood, -20 ℃ Cryopreserved blood Samples
Genotyping of known SNPs (heterozygous A locus: FAM + /Vic - , heterozygous B site: FAM + /Vic - , heterozygous C site: FAM + /Vic - ) was used to perform genotyping test on fresh blood sample A, and the test result was accurate (Fig.2).
Fig.2 Detection results of different SNP sites after lysis and release of fresh blood sample A. The blood was lysed and centrifuged to obtain the supernatant, and 4 μL of the supernatant was added to the 20 μL reaction system as a template for qPCR. The test results were consistent with the Sanger sequencing results.
Genotyping of known SNPs (heterozygous A locus: FAM + /Vic - , heterozygous B site: FAM + /Vic - , heterozygous C site: FAM + /Vic - The refrigerated blood sample A (stored at 4℃ for 2 months) was used for genotyping detection using this kit, and the test results were accurate (Fig.3).
Fig.3 Detection results of different SNP sites after lysis and release of stored blood sample A. The blood was lysed and centrifuged to obtain the supernatant, and 4 μL of the supernatant was added to the 20 μL reaction system as a template for qPCR. The test results were consistent with the Sanger sequencing results.
- Nucleic acid release detection results fluorescence signal intensity and Ct Value comparable to high purity obtained by nucleic acid extraction DNA Amplification results
200 μL EDTA blood was taken from each sample A and treated with the nucleic acid release agent in this kit and the whole blood genomic DNA extraction kit (FS-B8001) respectively. When the same supernatant/elution volume was obtained, 4 μL of supernatant was taken as a template to detect the B site respectively. The results showed that the amplification results of the whole blood after simple treatment by this kit were consistent with the Ct value of the whole blood nucleic acid extraction and purification test, and the fluorescence intensity remained unchanged (Fig. 4).
Fig.4 Comparison between Qihengxing blood direct amplification kit and blood nucleic acid extraction kit. The thick line is the qPCR result after nucleic acid release solution treatment, and the thin line is the qPCR result after whole blood genomic DNA extraction and purification. The blue is the FAM channel, the orange is the Rox channel (internal reference), and the green is the Vic channel.
Product details consultation/product trial application: 010-62149251/15801217100 (same number on WeChat)
