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低温
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- 供应商:
上海圻明生物
- 规格:
50次
Rhipicephalus spp.棕蜱染料法荧光定量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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文献和实验处于蜱类环境中的哺乳动物。在我们的研究中,我们在华盛顿州切尼的特恩布尔国家野生动物保护区(TNWR)调查了花栗鼠和鹿鼠等小型哺乳动物的立克次体抗体。我们的程序是从这些小型哺乳动物身上采集血样,并使用斑点热群立克次体EIA-IgG抗体试剂盒中含有抗体的血清。这个试剂盒有小井,里面有立克次体的抗原,所以如果血清中有这种疾病的抗体,抗体会粘在抗原上,我们会有一个阳性的测试结果,导致试剂盒运行时的颜色变化。如果阳性,这可能意味着这些动物被引入立克次体细菌,这种病原体在我们地区很普遍。在我们的研究中
,不靠壁,不贴液面。 混匀平均分装的注意事项: 1、第一管吸液前需要先把枪头在液面中浸润一下,以保证三复孔的枪头吸液量是差不多的。 2、用混匀器进行混匀,不要用移液器吹打。 详细说明书 transhold cat. A2010A0112荧光定量PCR Premix试剂盒(荧光染料) (FQ
所特有的,下游引物为通用引物就可以了。 荧光定量PCR检测方法有SYBR Green染料法和TaqMan探针法。前者需要调整引物浓度以及引物扩增效率,把引物二聚体调整到越小越好;探针法则需要设计荧光探针,这其中由于MGB探针需要碱基数量少和特异性好的特点而被推荐。 探针设计位置有3个:完全与miRNA序列相同、在miRNA与RT引物的交叉点、完全在RT引物上;这其中又有正向和反向互补两种情况。至于探针要设计在那个位置,根据自己试验情况而定,本人以为效果都差不多。 四、试验操作流程
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