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- 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:
2μg
鼻疽假单胞菌PCR阳性对照质粒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,在实际应用中受到限制。PCR 在结核菌检测方面有简便、敏感、特异的优点。一般认为样本中内要有100个左右的结核菌即可被检出。目前用于结核菌PCR 诊断的试剂,其引物主要来源于以下基因片段36KD/65KD抗原蛋白基因;染色体重复插入序列IS986、IS960、IS6110、染色体质粒DNA PH7311、PMTB4、P36基因等。其中最常用的是染色体重复插入序列IS986或IS6110,1990年Hermans首先介绍并使用了IS986基因设计的引物扩增产物为245BP,研究表明这一基因对人型
察结果。 (3)结果:出现红色为阳性反应。如欲观察有无氮气产生,可于培养基管内加1只小倒管,有气泡产生,则表示有氮气生成。如欲检查培养基中硝酸盐是否被分解,可取锌粉少许加入培养基内,如出现红色表明硝酸盐仍存在;若不出现红色,表示硝酸盐已被分解。 (4)应用:本试验广泛用于细菌鉴定。肠杆菌科细菌均能还原硝酸盐为亚硝酸盐;假单胞菌属中有的细菌能产生氮气,如铜绿假单胞菌、嗜麦芽窄食单胞菌、斯氏假单胞菌,有的则能还原硝酸盐为亚硝酸盐,如鼻疽假单胞菌等;厌氧菌如韦荣菌也能还原硝酸盐为亚硝酸
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