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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:
2μg
液化沙雷菌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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文献和实验保存的菌种,在试验前必须进行菌株的生物特性鉴定。菌株鉴定的判断标准,如表1所示。表1 试验菌株鉴定的判断标准菌株组氨酸缺陷脂多糖屏障缺损氨苄青霉素抗性切除修复缺损四环素抗性自发回变菌落数*TA97TA98TA100TA102+++++++++++++++----+90-18030-50100-200240-320注「+」表示需要组氨酸「+」表示具有 rfa 突变「+」表示具有 R 因子「+」表示具有 △uvrB 突变「+」表示具有 pAQ1 质粒*在体外代谢活化条件下自发回变菌落数略增 7.2
,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
的胞外蛋白水解酶(明胶酶)能分解明胶,使明胶失去凝固能力而液化。 (2)方法:将待检菌接种于明胶培养基中,35℃孵育24h到7d或更长时间,每24h取出放入4℃冰箱约2h后,观察有无凝固。 (3)结果:如无凝固,则表示明胶已被水解,液化试验阳性。如凝固,则继续培养。 (3)应用:奇异变形杆菌、普通变形杆菌、沙雷菌属和阴沟肠杆菌等到能液化明胶,肠杆菌科中的其它细菌很少液化明胶。有些厌氧菌如产气荚膜梭菌、脆弱类杆菌等也能液化明胶。另外,许多假单胞菌也能产生明胶酶而使明胶液化。 3.
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