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上海圻明生物
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Shigella spp.志贺氏菌属探针法荧光定量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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文献和实验志贺氏菌属( shigella )是一类革兰氏阴性杆菌,是人类细菌性痢疾最为常见的病原菌,通称痢疾杆菌。大小为 0.5 ~ 0.7 × 2 ~ 3 μ m ,无芽胞,无荚膜,无鞭毛。多数有菌毛。革兰氏阴性杆菌。兼性厌氧菌,能在普通培养基上生长,形成中等大小,半透明的光滑型菌落。在肠道杆菌选择性培养基上形成无色菌落。分解葡萄糖,产酸不产气。 vp 试验阴性,不分解尿素,不形成硫化氢,不能利用枸橼酸盐作为碳源。宋内氏志贺氏菌能迟缓发酵乳糖( 37 ℃ 3 ~
Polymerase Chain Reaction Detection of Invasive Shigella and Salmonella enterica in Food
The ability to detect Shigella in foods is often hampered by the often-low numbers of organisms present in the sample at the time of analysis. At present, a reliable, time-efficient, and highly sensitive protocol to isolate Shigella
相关专题 实验室很多同学都要做Real time PCR实验,实验室的师兄师姐都会有很多宝贵意见,不过也有实验室前没有做过的,查找了下资料和大家分享下关于实时荧光 Taqman 探针设计、实时荧光PCR探针的选择、引物的设计及评价。 荧光探针法是用序列特异的荧光标记探针来检测产物,探针法的出现使得定量PCR技术的特异性比常规PCR技术大大提高。目前较常提及的有TaqMan探针、FRET杂交 探针(荧光共振能量传递探针)和分子
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