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低温
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详见说明
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99
- 供应商:
上海圻明生物
- 规格:
2μg
豚鼠耳炎诺卡菌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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文献和实验诺卡菌属细胞壁含分枝菌酸,是广泛分布于土壤中的需氧性放线菌。多数为腐物寄生性的非病原,不属于人体正常菌群,故不呈内源性感染。分为星形诺卡菌、短链诺卡菌、鼻疽诺卡菌、肉色诺卡菌、巴西诺卡菌、越橘诺卡菌、豚鼠耳炎诺卡菌、南非诺卡菌、苦味诺卡菌等9种。对人致病的主要有3种:星形诺卡菌、豚鼠诺卡菌和巴西诺卡菌。引起人类疾病 主要为星形诺卡菌和巴西诺卡菌。在我国最常见的为星形诺苄菌。 奴卡菌病多为外源性感染,可因吸入肺部或侵入创口引起化脓感染。如星形诺卡菌主要通过呼吸道进入人体引起人的原发
批黧的螺旋质粒DNA,分装成许多小份贮存于-70℃。这些标准制品可用来检验每一批新的感受态细胞的转化效率,并检查每一个实验的转化效率。设立这样一个阳性对照后,如果某一次实验得不到转化菌落,就可以根据对照的情况查明宣究竟是感受态细菌方面有庇漏,还是DNA制品间有差异。分装的感受态细菌可在-70℃保存几个月而转效率无明显下降。1)用无菌铂丝直接蘸取冻存有大肠杆菌DHl株(或DH5株、MM249株原种)(贮存于-70℃的冻培养基上,见附录A),在SOB琼脂平板表面划线, 于37℃培养16小时。将冰冻
,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
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