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上海圻明生物
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SARS-CoV-2冠状病毒(新冠病毒)N基因探针法荧光定量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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文献和实验任何总RNA提取试剂在提取过程中无法完全避免DNA的微量残留,本公司的EASYspin系列RNA提取产品,由于采取了本公司独特的缓冲体系和选择了特殊吸附能力的吸附膜,在大多数RT-PCR 扩增过程中极其微量的DNA残留(一般电泳EB染色紫外灯下观察不可见)影响不是很大, 如果要进行严格的mRNA表达量分析如荧光定量PCR,我们建议在进行模板和引物的选择时:1) 选用跨内含子的引物,以穿过mRNA中的连接区,这样DNA就不能作为模板参与扩增反应。2) 选择基因组DNA和cDNA上扩增的产物
,在此基础上再进行PCR。由此可见,RT-PCR在基因表达研究和对RNA病毒的基因诊断中具有特殊作用。 (4)荧光定量PCR:本方法通过对照基因的扩增或对引物进行荧光标记,结合一定的仪器可以对PCR产物进行定量。 (5)联合PCR技术。 三、核酸序列分析法: 核酸序列分析法是最确切的基因诊断分析法,它通过测定碱基排列序列而发现DNA的具体变异情况。 核酸序列分析法有两种:化学裂解法和双脱氧核苷酸末端终止法。 基因诊断的应用 1.病原生物的侵入:一般侵入体内的病原生物可通过显微
-transferase)二、足印法(Footprinting) 足印法(Footprinting)是一种用来测定DNA-蛋白质专一性结合的方法,用于检测目的DNA序列与特定蛋白质的结合,也可展示蛋白质因子同特定DNA片段之间的结合。其原理为:DNA和蛋白质结合后,DNA与蛋白的结合区域不能被DNase(脱氧核糖核酸酶)分解,在对目的DNA序列进行检测时便出现了一段无DNA序列的空白区(即蛋白质结合区),从而了解与蛋白质结合部位的核苷酸数目及其核苷酸序列。三、染色质免疫共沉淀技术(Chromatin
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