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- 文献和实验
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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
亚巴猴肿瘤病毒病毒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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文献和实验过程中加样器应该专用,不能交叉使用,尤其是PCR产物分析所用加样器不能拿到其它两个区; 8、重复实验,验证结果,慎下结论。 二、 追踪污染源 如果不慎发生污染情况,应从下面几条出发,逐一分析,排除污染。 (一)设立阴阳性对照:有利于监测反应体系各成分的污染情况。选择阳性对照时,应选择扩增弱,且重复性好的样品,因强阳性对照可产生大量不必要的扩增序列,反而可能成为潜在的污染源。如果以含靶序列的重组质粒为对照,100个拷贝之内的靶序列就足以产生阳性扩增。阴性对照的选择亦要慎重,因为PCR敏感性极高
设计:客户提供靶基因名称、序列或GeneBank ID号 2) shRNA载体构建(靶基因4个,阴性对照1个,阳性对照1个) 3) 质粒扩增、纯化 4) 转染靶细胞(客户提供靶细胞及其培养方法等相关资料) 5) 抑制效果检测:实时荧光定量PCR检测mRNA表达,Western Blot检测蛋白表达 6) 提供实验报告:shRNA序列、测序报告、荧光定量PCR报告、WB报告等。 ■ 慢病毒介导的RNA干扰
浅析染色质免疫沉淀(ChIP)技术在 DNA 与蛋白质相互作用研究中的重要性
染色质免疫沉淀(ChIP)是研究蛋白质-DNA 相互作用的一项强大技术,广泛用于多个领域的染色质相关蛋白的研究(如组蛋白及其异构体,转录因子等),特别适用于已知启动子序列或整个基因位点的组蛋白修饰分析研究。这项技术采用特定抗体来富集存在组蛋白修饰或者转录调控的 DNA 片段,通过多种下游检测技术(定量 PCR ,芯片,测序等)来检测此富集片段的 DNA 序列。 ChIP 技术自诞生之后,已成功的应用于人或动物细胞和组织 [1] 、植物组织 [2] 、酵母 [3] 以及细菌、质粒
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