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Enterovirus(EV)肠道病毒C组探针法荧光定量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【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enterovirus 71(EV71) JOURNAL Unpublished REFERENCE 2 (bases 1 to 7414) AUTHORS Hu,A.-r., Gu,W.-z. and Jiang,S.-w. TITLE Direct Submission JOURNAL Submitted (03-DEC-2010) Science and Education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
普通感冒竟用基因编辑去攻克?Nature 子刊揭示 SETD3 基因与病毒增殖有关
,我们熟知的鼻病毒、冠状病毒及副流感病毒只是类别,细分的病毒株没法计数,而且随时有崭新病毒株产生。所有的这些都导致难以找到致病原的共性。这一次,为了攻克感冒这一「蕞尔小疾」,科学家选择动用「基因编辑」这门高射炮。这篇题为《Enterovirus pathogenesis requires the host methyltransferase SETD3》的研究文章发表在 9 月 16 日的 Nature 子刊上。图片来源:nature microbiology文章思路:第一步是慎重选择研究对象——研究
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