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Squama manis穿山甲探针法PCR鉴定试剂盒上海圻明生物优势供应。更多产品资料欢迎免费咨询。
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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文献和实验实时荧光定量 PCR 技术(Quantitative Real-time PCR,简称 qPCR)是在 PCR 扩增过程中,通过实时监测荧光信号的变化,达到对待检测样本中初始模板定量分析的方法。当前 qPCR 技术被广泛应用于临床疾病诊断,动物疾病监测,食品安全分析等领域。随着 2020 年新冠疫情在全球的蔓延,qPCR 技术更是因其检测通量高,速度快,操作简便等优势成为新冠病毒核酸检测的有效方法被众人所熟知。 那么为了得到可靠、准确的检测结果,我们需要注意哪些方面呢?其实决定一次实验成功
继管轶之后,华南农大论文上线!新型冠状病毒中间宿主溯源指向穿山甲
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2020 年 2 月 9 日,华中农业大学陈焕春教授在湖北省疫情防控工作新闻发布会表示,新冠病毒可能存在多个中间宿主。而早在 2 月 7 日时,华南农业大学发布消息,该单位沈永义教授、肖立华教授等的研究团队通过分析 1000 多份宏基因组样品,并结合其他研究结果后,确定穿山甲为新型冠状病毒的潜在中间宿主。图片来源:华南农业大学研究人员继而通过分子生物学检测,揭示穿山甲中 β 冠状病毒的阳性率为 70 %;进一步对病毒进行分离鉴定,电镜下观察到典型的冠状病毒颗粒结构。图片来源:华南农业大学最后
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