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- 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
猪轮状病毒D群RT-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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文献和实验为普通轮状病毒(主要引起婴幼儿腹泻);B群为猪轮状病毒和成人腹泻轮状病毒(adult diarrhea rotavirus),后者为我国学者从1984年流行于我国各地的成人流行性腹泻患者分离出的一种新轮状病毒,目前将此种病毒与世界其它地区发现的副轮状病毒、类轮状病毒统称为抗原特异性轮状病毒;C群为人和猪轮状病毒;D群为鸡和鸟类轮状病毒。成人腹泻轮状病毒与普通人轮状病毒在病毒抗原性、核酸图型及临床表现等方面均有差异。 轮状病毒对理化因素的抵抗力较强,耐乙醚和弱酸,在-20℃可以长期
)从倍增基因获得,也可以来自人工合成的基因。通过PCR获得感兴趣的DNA的方法是:首先,分离含有感兴趣的DNA片段的mRNA的总RNA,并通过逆转录聚合酶链反应(RT-PCR)将RNA逆转录成cDNA。根据公布的NCBI基因编码序列(CD)设计寡核苷酸引物,并在引物中加入指定的限制酶位点。然后,通过PCR扩增感兴趣的DNA的编码区,用凝胶提取试剂盒纯化后,将DNA片段插入克隆载体中。通过RT-PCR将RNA逆转录成cDNA通过PCR扩增感兴趣的DNA的编码区经验虽然从RT-PCR中获取DNA片段是常用
组DNA等可能的DNA污染;体外T7, T3, SP6等RNA Polymerases催化的RNA转录后去除DNA模板; DNase I footprinting研究DNA-蛋白质相互作用;缺口平移(nick translatioin);产生DNA随机片断文库;细胞凋亡TUNEL检测中部分剪切基因组DNA作为阳性对照。 来源:从牛胰腺纯化得到。 分子量:约32kDa(单体)。 活性定义:37℃10分钟内,将能够完全降解1μg pBR322质粒DNA所需的酶量定义
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