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Enterovirus(EV)肠道病毒染料法荧光定量RT-PCR试剂盒,停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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文献和实验分钟内快速纯化高品质DNA或者RNA ♦ 无需酚/氯仿抽提,无需乙醇沉淀 ♦ 重复性强,产量高,洗脱体积50-200 µl ♦ 纯化灵敏度高,含有carrier RNA组份,>80%回收率 ♦ 完全去除污染物和抑制剂,适用于各种下游应用\ 左图为肠道病毒EV71型提取核酸RNA后进行荧光定量PCR的检测结果,样本为肠道病毒EV71型灭活病毒培养液,病毒含量分别为5×106copies/ml、5×105copies/ml、5×104copies/ml
【求助】我在genebank提交核苷酸序列遇到困难,请求帮助
enterovirus 71(EV71) JOURNAL Unpublished REFERENCE 2 (bases 1 to 7414) AUTHORS Hu,A.-r., Gu,W.-z. and Jiang,S.-w. TITLE Direct Submission JOURNAL Submitted (03-DEC-2010) Science and Education Division, Ningbo Infectious Diseases
Transcriptome Profiling of HostMicrobe Interactions by Differential Display RT-PCR
cells to infection with a neurovirulent strain of enterovirus 71 (EV71) at different time-points during the infection process compared with uninfected cells. Using differential display RT-PCR, we identified mRNAs that were up- or down-regulated
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