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低温
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- 供应商:
上海圻明生物
- 规格:
50次
Beauveria bassiana球孢白僵菌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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文献和实验球孢白僵菌Beauveria bassiana (Bals.) Vuill 属于链孢霉科。寄生于家蚕幼虫体内(可寄生于60多种昆虫体上),使家蚕病死。干燥后的尸体称为僵蚕。入药能祛风、镇惊等。由于加强防治。近年来白僵菌对家蚕的感染大为减少,为解决僵蚕的药源问题,以蚕蛹为原料,接入白僵菌,所得蚕蛹可代僵蚕用。
是接头引物处于“Y”接头的2个分叉单链上,序列与接头一样,只有与特异引物引导合成了接头的互补序列后,接头引物才能退火参与扩增,流程如图4。 方卫国等尝试将YADE法引入到昆虫病原真菌的分子生物学研究,并取得了成功,建立了适合于球孢白僵菌和金龟子绿僵菌YADE体系。在已克隆的类球孢白僵菌类枯草杆菌蛋白酶基因CDEP-1的基础上,利用YADE法,克隆到该基因的启动子CDEPP。 先酶切球孢白僵菌基因组DNA ,然后与“Y”形接头相连,取连接产物做模板,先以基因
与接头一样,只有与特异引物引导合成了接头的互补序列后,接头引物才能退火参与扩增,流程如图4。 方卫国等尝试将YADE法引入到昆虫病原真菌的分子生物学研究,并取得了成功,建立了适合于球孢白僵菌和金龟子绿僵菌YADE体系。在已克隆的类球孢白僵菌类枯草杆菌蛋白酶基因CDEP-1的基础上,利用YADE法,克隆到该基因的启动子CDEPP。 先酶切球孢白僵菌基因组 DNA,然后与“Y”形接头相连,取连接产物做模板,先以基因特异引物1做线性扩增,再以线性扩增产物为模板,以接头引物和基因特异引物2做指数
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