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低温
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- 供应商:
上海圻明生物
- 规格:
50次
Radix stemonae百部PCR鉴定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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文献和实验的,为此政府已建立了检测食品中转基因成分的体制。其中,最重要的是转基因检测方法的可靠性和有效性。所以我们利用分子生物学技术建立起一套基于PCR反应的实验方案,用于食品质控实验室的工作。最要害的是第一步:DNA提取,我们做了很严格的监控 。我们依据植物基因组和质粒DNA的提取效率来选择DNA提取试剂盒。整个提取过程包括破坏细胞壁、去除RNA和利用沉淀去除蛋白质,然后将DNA吸附在层析柱上,经过洗涤后再洗脱下来。类似的方法已经用于从其他一些植物和病毒中提取DNA。PCR反应的过程分为三个阶段:(1)通过加热
。 PCR扩增每个基因座的多态性区域是分析精子的第一步。两个基因座用两组引 物,同时引物必须位于多态性区域的边侧。当精子两个靶序列经PCR扩增,就确定了 每个基因座的等位基因成份。人工合成的寡核苷酸探针可识别等位基因小到仅发生单 个碱基置换的实验方法已得到改进,这些等位基因牧场划的寡聚物(ASO)探针第一 次应用是将人正常β珠蛋白基因中的正常βA等位基因与镰刀形红细胞(βA)的突变 区分开来。本方法检测等位基因需要人工合成的小片段寡核苷酸(典型的长19bp)。 每个寡核苷酸与一个等位
min左右后进行电泳。(3)溴化乙锭(EB)(10mg/ml):在20ml水中加入0.2g溴化乙锭,磁力搅拌数小时。然后用铝箔包裹容器或将溶液转移至棕色瓶中,室温保存。(4)加样缓冲液(Loading Buffer)一般为6×Loading Buffer二、电泳鉴定时注意事项:佩戴一次性手套,避免皮肤接触EB。无路做胶还是电泳缓冲液尽量用新的,不要超过两周。三、电泳步骤1、50×TAE取10ml稀释成500ml,取50ml溶解0.5g琼脂糖。电炉上煮沸后加入EB 5ul。另外450ml TAE倒入
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