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Simian Virus 5 (SV5)猴病毒5 染料法荧光定量RT-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,SVDV和VSV特异性扩增的3对引物,并通过对扩增体系和扩增条件的优化,建立了特异、敏感、快速的能同时检测FMDV,SVDV和VSV的多重RT-PCR方法。 1 材料与方法 1.1 病毒株 猪水疱病病毒RNA、标淮阳性血清、标淮阴性血清和参考样品由英国动物卫生研究所Pirbright实验室惠赠;VSV-NJ病毒株和VSV-IND病毒株从美国国家兽医服务实验室(NVSL)引进,由本实验室保存;FMDV O疫苗株从云南保山疫苗厂引进,由本实验室保存。 1.2 试剂
immunodeficiency virus) 、 SIV (Simian immunodeficiency virus) 。其中研究最多最为透彻的是HIV。 重组 慢病毒的产生:瞬时转染法,即将包装结构和载体结构瞬时共转染法如293T 高表达细胞系而产生重组慢病毒。此法非常成功,大多实验室采用此法。包膜质粒、包装质粒与载体质粒共转(多用磷酸钙共沉淀法)染293T细胞直接产生生产细胞。最后重组慢病毒分泌到培养基中进行培养而得到大量载体慢病毒。 三质粒来包装产生重组 慢病毒
猿类病毒 为猿类病毒 simian virus 40 的缩写,亦称空泡化因子( vacuolating agent )属于乳头多瘤空泡病毒种( papovavious ),多型瘤病毒属。( polyomavirus )。由 B . H . Sweet 和 M . R . Hilleman ( 1960 )从猕猴( Macaca mulatta )中分离获得。 B . E . Eddy ( 1962 )和 A . Girardi ( 1962 )等确认 Sv 40 病毒
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