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上海圻明生物
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猕猴源性成分染料法荧光定量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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文献和实验标记(生物素、荧光、地高辛和Eu3+等)、引入与蛋白质结合的DNA序列、引入突变位点、插入与缺失突变序列和引入启动子序列等。 2 特定PCR的引物设计 在尽量遵循引物设计基本原则的同时,根据不同的实验目的,需要注意一些相应的事项,总结如下: 01 荧光定量PCR 荧光定量PCR有染料法和探针法两种。染料法只需要设计引物,而探针法除设计引物之外还得设计一条探针。 引物设计要尽量满足以下要求: 确保模板是cDNA
如果您的样本是组织,取样的过程非常重要。组织内部是有内源性 RNase 存在的,组织离体后内源性 RNase 则开始发挥作用,RNA 产生降解。所以离体的组织必须立马放在液氮中速冻,之后转移至 -80℃ 长期保存,且避免反复冻融。研磨组织的过程中需要保证在液氮环境中研磨。RNA 提取的过程中,保证样本的上样量不超过提取试剂盒说明书中规定的最大上样量,上样量过多,同样会造成 RNA 降解。 ②如果您的样本是细胞,则样本搜集过程要简单的多了,只要保证细胞状态良好(细胞状态差也容易造成提取的 RNA 降解
酶及其作用 (5)主要实验流程 2.组织类型决定制备方案的差异 (1)方案差异化的决定因素 细胞外基质(ECM)的成分与密度:例如,纤维化肿瘤的胶原蛋白含量远高于脑组织。 细胞间连接的紧密程度:上皮组织的紧密连接比淋巴组织要牢固得多。 组织内源性酶的含量:胰腺等组织富含内源性蛋白酶,解离时易发生自溶。 目标细胞类型及其敏感性:神经元等细胞对酶解和机械力非常敏感,而肿瘤细胞通常较为耐受。 (2)一张表快速了解不同组织类型 神经组织的单细胞制备是所有组织类型中最具挑战性的领域
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