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低温
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详见说明
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99
- 供应商:
上海圻明生物
- 规格:
2μg
猴D型逆转录病毒RT-PCR阳性对照质粒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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文献和实验体细胞选出;④具有促进外源DNA表达的调控区。重组DNA技术中最常用的载体有质粒、噬菌体λ,柯斯质粒(cosmid)和噬菌体M13。它们的受体细胞都是大肠杆菌。这四种载体的大小和结构尽管各不相同,但它们的共同特点是:①都能在大肠杆菌中自主复制,而且能连同所带的外源DNA一起复制;②都很容易同细菌DNA分开并加以纯化;③都有一段DNA对于它们自身在细菌中的增殖不是必需的。因此,外源DNA可以插入这一段DNA中,或是置换这一段DNA而不影响载体的复制。根据这一特点,载体又可分成插入型和置换型两大类。一、质粒
1、慢病毒载体概况 慢病毒是逆转录病毒科亚科之一,分为两类:灵长类和非灵长类慢病毒。灵长类慢病毒包括HIV-1、HIV-2、猴免疫缺陷病毒( SIV),非灵长类慢病毒包括猫免疫缺陷病毒(FIV)、牛免疫缺陷病毒(BIV)、马免疫缺陷病毒(EIAV)等。 多项研究表明慢病毒载体具有可感染非分裂期细胞、可长期表达、而且宿主细胞免疫反应小、细胞毒性小等优点。因其自身结构特点,可包装8~lOkb,适用于多基因表达系统,而且基因组包含较少的核苷酸,抵抗基因沉默能力较强,宿主体内更以
麻痹而死亡。 4猴逆转录病毒包括4种病毒:猴逆转D型病毒(Simian retrovirus D, SRV)、猴免疫缺陷病毒( Simian immunodeficiency virus, SIV)、猴T细胞趋向性病毒1型(Simian T lymphotropic, STLV-1)、泡沫病毒(Simian foamy virus,SFV)。
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