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上海圻明生物
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50次
Malleomyces mallei马鼻疽杆菌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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文献和实验补体结合反应技术(Complement fixation reaction technique)概
结合反应是诊断人、畜传染病常用的血清学诊断方法之一。本法不仅可用于诊断传染病,如鼻疽、牛肺疫、马传染性贫血、乙型脑炎、布氏杆菌病、钩端螺旋体病、血锥虫病等,也可用于鉴定病原体,如对马流行性乙型脑炎病毒的鉴定和口蹄疫病毒的定型等。 上一篇:经典补体结合反应(以马鼻疽补反为例) 下一篇:间接凝集反应技术(Indirect agglutination reaction technique)
,如破伤风杆菌,产气荚膜杆菌,炭疽杆菌,鼠疫杆菌等侦检. ⑤需特殊培养的无芽胞厌氧菌,如脆弱类杆菌、艰难杆菌的鉴定等. 某些病原微生物,某些遗传病或癌基因,型别较多,或突变或缺失存在多个好发部位,多重PCR可提高其检出率并同时鉴定其型别及突变等可系统应用的有:乙型肝炎病毒的分型;乳头瘤病毒的分型;单纯疱疹病毒的分型;杜氏肌营养不良症的分型及癌基因的检测等. 多重PCR的特点有: ①高效性,在同一PCR反应管内同时检出多种病原
。 (一)病原体测定 由于PCR技术的问世,使得病原体检测能够快速而方便的进行。但由于其高灵敏性,实验操作很容易受到污染而出现假阳性。只要有微量病原体存在,PCR扩增即可为阳性结果,但并不能作为诊断依据,只有当一定数量的病原体存在时才有临床意义,因此结模板定量显得特别重要。常规PCR由于不能定量而限制了其应用,然而,PE公司研制的FQ-PCR技术给解决这一问题提供了可能。目前该技术已经应用于丙肝病毒、人类乳头瘤病毒、结核杆菌和食品中大肠杆菌等许多病原体的检测研究。 Morris等[3]用FQ-PCR
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