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羊溶血性曼氏杆菌探针法PCR检测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,通过结合有链霉亲和素的磁珠对特异性标记有生物素的靶标序列进行纯化,大大提高了特异性。 武汉中帜生物携手美国Signosis,为广大科研用户提供特异性更高、分辨率更强的miRNA实时荧光定量PCR检测试剂。
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
不过科研人员就没那么好运了,科研中需要用到定量PCR检测的目标广泛,不容易找到带有探针的定量检测试剂盒。当然,针对这种现状,几个大公司陆续推出探针库,比如罗氏的UPL通用探针库,就可以通过组合定量检测出人、大小鼠等7个物种的大部分ORF序列,特别适合大型实验室;ABI公司现在可以根据用户的需要提供约十二万种基因表达引物、探针和两百多万种SNP分析的引物探针进行科学研究,据说是源自于世界各地研究人员要求定制探针或者引物后汇总的结果,可惜就是贵,就连美国的实验室教授
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