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上海圻明生物
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50次
Adenovirus(AV)腺病毒A型染料法荧光定量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(1)原理 腺病毒(adenovirus,AV)是一种没有包膜的线状双链DNA,它能感染分裂细胞和非分裂细胞,在核内以神经元细胞的形式复制,存在多种AV血清型。到目前为止,构建的绝大多数载体都是基于血清型2和5,通过转基因的方法取代E1或E3基因,降低病毒的复制能力。这些重组病毒仅在表达高水平E1和E3基因的细胞中复制,因此适合应用于治疗的高效控制系统。腺病毒载体能够高效传递和表达基因的能力(尤其是在体外),由于具有宿主范围广,对人致病性低;在增殖和非增殖细胞中感染和表达基因;能有效
(1)原理 腺病毒(adenovirus,AV)是一种没有包膜的线状双链DNA,它能感染分裂细胞和非分裂细胞,在核内以神经元细胞的形式复制,存在多种AV血清型。到目前为止,构建的绝大多数载体都是基于血清型2和5,通过转基因的方法取代E1或E3基因,降低病毒的复制能力。这些重组病毒仅在表达高水平E1和E3基因的细胞中复制,因此适合应用于治疗的高效控制系统。腺病毒载体能够高效传递和表达基因的能力(尤其是在体外),由于具有宿主范围广,对人致病性低;在增殖和非增殖细胞中感染和表达基因;能有效
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