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上海圻明生物
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50次
Hyaloma spp.眼蜱染料法荧光定量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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文献和实验的检测通道是只为自已厂家的专用的荧光染料或试剂开放的,有效检测通道也绝非像宣传资料中宣称的那么多。在购买前确认荧光定量PCR仪器的有效检测通道尤为重要,不能单单只听宣传。 考虑多重荧光定量PCR的通道数的时候也应该从实验室实际情况出发,多重PCR并非人人适用、人人可用,因为它使实验复杂化了。 误区二:real-time PCR仪无需梯度功能 对于使用染料法的定量PCR反应,虽然有各种各样的PCR引物设计软件或者经验公式计算熔解温度(Tm值),但运用的公式不同、引物序列不同
的检测通道是只为自已厂家的专用的荧光染料或试剂开放的,有效检测通道也绝非像宣传资料中宣称的那么多。在购买前确认荧光定量PCR仪器的有效检测通道尤为重要,不能单单只听宣传。 考虑多重荧光定量PCR的通道数的时候也应该从实验室实际情况出发,多重PCR并非人人适用、人人可用,因为它使实验复杂化了。 误区二:real-time PCR仪无需梯度功能 对于使用染料法的定量PCR反应,虽然有各种各样的PCR引物设计软件或者经验公式计算熔解温度(Tm值),但运用的公式不同、引物序列不同
,根据其更换宿主的次数可分为四种类型:①单宿主蜱:发育各期都在一个宿主体上,雌虫饱血后落地产卵。如微小牛蜱(Boophilus microplus)。②二宿主蜱:幼虫发育为若虫在一个宿主体上,而成虫在另一个宿主体上寄生。如残缘璃眼蜱(Hyaloma detritum)。③三宿主蜱:幼虫、若虫、成虫分别在3个宿主体上寄生。如全沟硬蜱、草原革蜱。90%以上的硬蜱为三宿主蜱,蜱媒疾病的重要媒介大多是三宿主蜱(图21-7)。④多宿主蜱:幼虫、各龄若虫和成虫以及雌蜱每次产卵前都需寻找宿主寄生吸血,每次饱血后离去
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