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低温
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- 供应商:
上海圻明生物
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50次
Flos lonicerae金银花探针法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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文献和实验实时荧光定量 PCR 技术(Quantitative Real-time PCR,简称 qPCR)是在 PCR 扩增过程中,通过实时监测荧光信号的变化,达到对待检测样本中初始模板定量分析的方法。当前 qPCR 技术被广泛应用于临床疾病诊断,动物疾病监测,食品安全分析等领域。随着 2020 年新冠疫情在全球的蔓延,qPCR 技术更是因其检测通量高,速度快,操作简便等优势成为新冠病毒核酸检测的有效方法被众人所熟知。 那么为了得到可靠、准确的检测结果,我们需要注意哪些方面呢?其实决定一次实验成功
高通量、低成本SNP、突变或甲基化检测方法—HRM 技术应用
细胞中1 个突变细胞,适用于手术和其它微量组织中突变检测。检测灵敏性远高于“PCR+ 测序”的25% ,即100 个正常细胞中至少有25 个突变细胞,测序仅适用手术组织。 • 特异性好:PCR 产物无需后续处理,特异性高达100% 。直接未知杂合突变鉴定准确性100%,特异性100% 。直接未知纯合突变鉴定准确性96%, 利用非标记探针快速基因分型法直接未知纯合子鉴定准确性100%。 • 重复性好:重复性100% • 使用范围广:不受碱基位点局限,除可检出已知突变外,也能检测出未知突变。
— real-time PCR,针对miRNA的real-time PCR与常规的real-time 有所不同。 目前有的公司提供的是探针法试剂,有的是SYBR Green法试剂,因探针法试剂成本较高,丁香通在此挑选出SYBR Green法的代表作 — QIAGEN公司的完整的一套miRNA定量试剂miScrip system来介绍,这次比起其他进口品牌同系列的产品,QIAGEN价格倒是不贵。 第一步是miRNA的提取,核酸提取无疑是QIAGEN公司的绝活,对于miRNA这样的小分
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