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低温
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详见说明
- 库存:
99
- 供应商:
上海圻明生物
- 规格:
2μg
猪霍乱沙门氏菌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S9 或未加 S9 条件下为阳性,均可报告该受试物对鼠伤寒沙门氏菌为致突变阳性。如果受试物经四个试验菌株检测后,无论加 S9 和未加 S9 均为阴性,则可报告该受试物为致突变阴性。 13 试验报告试验报告应包括以下内容:(1)受试物名称、理化性状、配制方法、使用溶剂;(2)试验菌株:所用试验菌株;(3)代谢活化系统:所用诱导剂;(4)试验方法:简述操作步骤,除受试物剂量分组外,还应说明空白对照、溶剂对照和阳性对照, 阳性结果判定标准;(5)结果:以列表方式报告受试物的 Ames 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
肉汤培养基中,于35℃孵育,观察2~8d。应同时做阴性对照。 (3)结果:紫红色为阳性,与对照管颜色相同为阴性。 (4)应用:主要用于沙门菌属内各菌种间的鉴别。伤寒沙门菌、甲(丙)型副伤寒沙门菌、猪霍乱沙门菌、孔道夫沙门菌和仙台沙门菌本试验为阴性,乙型副伤寒沙门菌结果不定,其它不常见沙门菌多数为阳性。 9.葡萄糖酸氧化试验 (1)原理:某些细菌可氧化葡萄糖酸钾,生成α-酮基葡萄糖酸。α-酮基葡萄糖酸是一种还原性物质,可与班氏试剂起反应,出现棕色或砖红色的氧化亚铜沉淀。 (2)方法
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