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上海圻明生物
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甲型流感(禽流感)H1N1/H3亚型双重RT-PCR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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文献和实验实验室检测和快速诊断的标准如《GB/T 19438.1-2004禽流感病毒通用荧光RT-PCR检测方法》、《GB/T 19438.2-2004 H5亚型禽流感病毒荧光RT-PCR检测方法》、《GB/T 19438.3-2004 H7亚型禽流感病毒荧光RT-PCR检测方法》等。以上实时荧光定量PCR技术在禽流感检测方面的国家标准是该技术在当前的人感染猪流感诊断中应用的基础和前提。实时荧光定量PCR技术及在我国卫生应急中的应用要求实时荧光定量PCR技术(Real-time quantitative
体,容易引起世界性大流行。由于病毒多变异,导致甲型流感反复发生,难以彻底根除。 结构、形状和化学组成 禽流感病毒基因组由8个负链的单链RNA片段组成。这8个片段编码10个病毒蛋白,其中8个是病毒粒子的组成成分(HA、NA、NP、M1、M2、PB1、PB2和PA),另两个是分子质量最小的RNA片段,编码两个非结构蛋白——NS1和NS2。NS1与胞浆包含体有关,但对NS1和NS2的功能目前尚不清楚。现在已经获得了包括H3、H5和H7在内的几个禽流感病毒亚型HA基因的全部序列
人:郭元吉、王敏,联系电话:010-63534632,010-63581337,传真:(010)63534632。 五、标本检测方法 1.禽H5 病毒核酸检测 材料:疑似禽流感患者的咽、鼻拭子或含漱液,死亡病例的尸检肺组织、气管分泌物。 方法: RT-PCR,用于禽H5 亚型流感病毒的快速检测、测序及排除工作 试剂:特异性H5 及甲型流感病毒的引物、病毒RNA 提取试剂盒、逆转录酶、Taq DNA 聚合酶等试剂, 建议使用德国QIAGEN 的Rneasy Mini Kit (病毒
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