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上海圻明生物
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Potato spindle tuber viroid(PSTVd)马铃薯纺锤块茎类病毒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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文献和实验结构比病毒更简单的新型植物病原体。是T. O Diener( 1971)在马铃薯瘦弱块茎病( potatospindle tuber disease)的病株上发现的,并予以命名。此外,已知的还有柑桔裂皮病( citrus exoc- ortis disease)、酒花麻矮化病( hop stunt dise- ase)等数种类病毒。这些都是由 1.1— 1.3× 105 道尔顿的一条小的 RNA所组成。在电子显微镜下可见到是与双链 RNA粗细相等的 50nm长的杆状分子,因为互补
病毒是复杂生命体系的最低极限。� 类病毒是一类能感染某些植物致病的单链闭合环状的 RNA 分子。类病毒基因组小,分子量为 1 × 10 5 。目前已测序的类病毒变异株有 100 多个,其 RNA 分子呈棒状结构,由一些碱基配对的双链区和不配对的单链环状区相间排列而成。它们一个共同特点就是在二级结构分子中央处有一段保守区。类病毒通常 246~399 个核苷酸。如马铃薯纺锤块茎类病毒 ( Potato spindle tuber viroid , PSTVd , Vd 是用来与病毒加以区别
植物RNA提取--Methods for Plant RNA Isolation
in a Sorvall ss-34 for 10min. Total is contained in the upper aqueous layer. This protocol works well for us. We use it to extract total for analysis with DIG-labelled c probe for PSTVd (Potato Spindle Tuber Viroid).
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