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Avian Nephritis Virus(ANV)禽肾炎病毒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-competitive protein binding 放免-竞争性蛋白结合 RANA rheumatoid anti-nuclea antigen 类风湿性关节 炎抗核抗原 RPGN rapid progressive nephritis 进行性肾小管肾炎 RPM rolls per minute 每分钟转速 RRA radioreceptoassay 放射受体分析 RT-PCR reverse transiption-PCR 反转录-PCR RSA
除净,与RNA结合后会影响逆转录和PCR;残存的RNase极易将膜板RNA降解掉。硫氰酸胍(GaSCN)-CsCl法或酸性硫氰酸胍-酚-氯仿法可提得理想的RNA制品,尤以后者方法为佳,适合一般实验室进行。 常用的逆转录酶有两种,即禽类成髓细胞性白血病病毒(Avian myeloblastosis virus, AMV)和莫洛尼鼠类白血病病毒(Moloney murine leukemia virus, MO-MLV)的逆转录酶(RT)。一般情况下用Mo-MLV-RT较多,但模板RNA的二级
rapid progressive nephritis 进行性肾小管肾炎 RPM rolls per minute 每分钟转速 RRA radioreceptoassay 放射受体分析 RT-PCR reverse transiption-PCR
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