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One of the many important uses of PCR technology is that it can be used to label DNA probes with high specific activity. PCR technology has high specificity, and can synthesize probe DNA fragments in quantities within 1~2h if [α-32P]dNTP or other markers are added to the substrate
dNTPs, the probe DNA can be well labeled during DNA synthesis, and the incorporation rate of the marker can be as high as 70%~80%. Therefore, PCR labeling technology is particularly suitable for large-scale detection and non-radiolabeling. The disadvantage of this method is that a specific pair of PCR primers is synthesized.
Labeling can also be achieved by using small fragments prepared from probe DNA as primers.
Solution preparation
1. Prepare a stock solution
Unless otherwise stated, all unused stock solutions should be divided into disposable aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
1.1* Acid Stock Solution (125X):
Add 20 μL DMSO to *ate (component B) to make a 125X* acid stock solution.
2. Prepare standard solutions
*Salt standard solution
Add 50 μL of 1 mM KH2PO4 (Component C) to 950 μL of deionized water or enzyme reaction buffer to give a 50 μM * saline standard solution (PS7). A 50 μM * saline standard solution (PS7) was taken and serially diluted 1:2 to obtain a serially diluted phosphate standard with deionized water or enzyme reaction buffer.
3. Prepare a working solution
Add 20 μL of 125X* stock solution to 2.5 mL of sterile H2O and mix well to make a working solution of *salt. Avoid potential Pi contamination. Note: Avoid direct exposure of *salts (component B) to light. Due to the high sensitivity of this assay to Pi, it is extremely important to use Pi-free labware and reagents.
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文献和实验and Parvovirus B19 viral antigen can also be detected in serum and plasma using antigen-specific ELISA. NAT systems have also been described to detect newly identified parvoviruses such as human bocavirus (HBoV), PARV4, and PARV5, although absolute confirmation
Green染料法的qPCR检测系统,可用于从一个合成反应的cDNA中检测多种miRNAs,不仅减少了误差、节约了样品,同时还实现了检测的高通2. 检测的特异性:独特的miRNA引物设计技术及其优化的反应体系避免了非特异性扩增、特别是Pre-miRNA的污染3.检测分辨率高:该系统能分辨单碱基差异的miRNA4.一站式解决方案:公司不仅提供检测的试剂盒,还提供经验证的人源、小鼠源、大鼠源的特异性miRNA 检测引物。5.检测灵敏度高:可在低至pg级的总RNA中检测到特异表达的miRNA参考文献:[1]
.org/cgi/content/abstract/276/5316/1268 另Nature上的此文,也被引用了1300次之多,足可借鉴: Systematic variation in gene expression patterns in human cancer cell lines http://www.princeton.edu/genomics/botstein/publications/2000_Ross_Systematic.pdf
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