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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
低温
- 保质期:
详见说明
- 库存:
99
- 供应商:
上海圻明生物
- 规格:
50次
Mannheimia varigena异源曼氏杆菌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(1 ,2 ) while A. suis, H. somnus , and Mannheimia hemolytica are considered as potential animal pathogens. The pathogenic potential is incompletely known for most taxa, probably due to limitations in their classification and identification, and inappropriate detection
互补。 第二次PCR的第一循环中,PCR产物与质粒均变性与复性。除自身复 性产物(这种复性产物不被扩增)外,PCR产物与质粒可通过各自3’ 端互补序列杂交成部分异源双链,延伸时,重叠的3’端互为此物沿 各自互补链延伸,结果可产生PCR片段与线性质粒的“连接”。然后 两管中PCR扩增各进行15―20个循环。这便可产生大量一端管1)或 另一端连接有PCR插入片段的质粒。 第二次PCR后,将第1管与第2管反应液混合,用碱变性双链,中和后 稀释变性的DNA。反应管中的单链DNA可以复性
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