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低温
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- 供应商:
上海圻明生物
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50次
Clostridium tetani破伤风梭状芽孢杆菌染料法荧光定量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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文献和实验究假破伤风梭状芽孢杆菌( Clostridium tetanomor-phum )的谷氨酸代谢过程中,通过确定对光不稳定的未知维生素 B 12 是辅酶这一事实后才发现的( 1958 )。现在知道除了这种谷氨酸变位酶外,还有作为参与甲基丙二酸单酰 CoA 的变构反应和蛋氨酸合成反应等十余种酶的辅酶。在自然条件下, B 12 在生物体内几乎全以这种形态存在,氰钴胺素是抽提过程中所得到的人工产物。而脱氧腺苷基是来自 ATP ,由 B 12 通过酶促作用生成的。
Clostridium difficile Toxinotyping
Clostridium difficile shows considerable variability in the PaLoc region encoding two main virulence factors, toxins TcdA and TcdB. Strains with changes in PaLoc are defined as variant toxinotypes and currently 27 such groups are recognized
Transposon Mutagenesis in Clostridium difficile
Genetic manipulation of Clostridium difficile is notoriously difficult, currently there is only one reliable method for generating random mutations in the organism and that is to use the conjugative transposon Tn916 . Tn916 enters the genome
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