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上海圻明生物
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小鼠线粒体DNA探针法荧光定量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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文献和实验上海西唐生物科技有限公司 021-55229872, 65333639 www.westang.com 小鼠 抗线粒体抗体(AMA) ELISA 试剂盒 ( 用于血清、血浆、细胞培养上清液和其它生物体液内 ) 原理 本实验采用双抗体夹心 ABC-ELISA 法。用抗小鼠 AMA 单抗包被于酶标板上,标准品和样品中的 AMA与单抗结合,加入生物素化的抗小鼠 AMA
actin,山东大学提供样品基因。cDNA制备:测定小鼠肝脏总RNA浓度后,使用BioTeke Super RT Kit 制备20μl cDNA.操作步骤参照试剂盒说明书。一步法荧光定量PCR:Bioteke One-step SYBR real-time RT-PCR Kit20μl反应体系包括:10μl 2×One-step SYBR premixture,7.2μl RNA Free Water,0.4μl Primer F,0.4μl Primer R,1μl cDNA,1μl One-step
迁移实验需要制胶、跑胶、转膜、检测等步骤,操作稍显繁琐;特别是在样本量较大的时候,更显得力不从心。 为了解决这些转录因子检测中的问题,美国Signosis开发出了一种快速高效的半定量活性转录因子检测方法——滤板法。这种方法同样基于转录因子与特定DNA序列结合的原理,以标准96孔板为载体进行实验操作。针对不同的转录因子设计特异性的生物素标记DNA探针,探针与样本核提取物中相应的转录因子结合形成TF/DNA复合物。用滤板除去游离的DNA探针,随后从滤板上洗脱并收集TF/DNA复合物中的DNA探针。用标准
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