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低温
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- 供应商:
上海圻明生物
- 规格:
2μg
小鼠白血病病毒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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基因编辑再次升级!领域大牛刘如谦 Cell 发文开发新工具,可安全高效进行体内基因编辑
了先导编辑(Prime Editor, PE)。刘如谦带领团队开发的这一系列基因编辑工具能够在不造成 DNA 双链断裂的情况下,实现对基因组的点突变进行定点矫正修复,因此被认为是比较安全的,也更具有临床应用前景。前期的研究工作也已经证实可以应用 BEs 来纠正小鼠和非人灵长类动物的致病点突变并改正疾病表型,强调体内碱基编辑作为一种治疗策略的潜力。 要想将碱基编辑技术广泛应用于临床治疗,那么就需要安全有效的方法将单碱基编辑器输送到多个组织和器官。截至目前,最常用和最有效的载体仍然涉及病毒的使用
liuruya 问别人要了个带GFP-tag(GFP-N2载体)的质粒,目的基因1.4kb,分子量约52kD,转染293T细胞能见荧光,WB杂不出来,设置了阴性和阳性对照证实不是WB的技术问题。后来又换了个flag-tag,仍然杂不出来WB。仔细检查过序列,不存在移码的问题,非常费解。有同学分析是空间位阻问题,不知换个tag是不是能解决这个问题。有同学遇到过这种问题么,或者帮忙分析一下可能的原因。谢谢!!! zhuqueleee
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