常见大肠杆菌种系 更多大肠杆菌信息可参考(https://blog.addgene.org/plasmids-101-common-lab-e-coli-strains?_ga=2.210903381.1761310573.1650342339-1554568351.1648354532) 2. 常用于质粒选择的抗生素 许多质粒被设计成包含抗生素抗性基因,当其表达时,仅允许含有质粒的细菌在含有该抗生素的培养基中或培养基上生长。这些...
启动子的强度对于控制插入片段的表达水平也很重要(即,强启动子指导高表达,而较弱的启动子可以指导低/内源表达水平)。有关细菌和真核生物启动子以及研究中使用的常见启动子的更多信息,请查看Addgene的启动子参考页面(https://www.addgene.org/mol-bio-reference/promoter-background/)。 Selectable Marker:选择性标...
由于质粒多聚化是由同源重组引起的，因此缺乏重组酶蛋白 RecA 的细菌菌株的质粒扩增可以降低多聚化频率，并且在某些情况下完全阻止质粒多聚体的形成。DH5α、HB101、NEB Stable、Stbl3等大肠杆菌菌株是recA突变菌株，具有更低的质粒多聚化频率。有研究表明，与DH5α相比，菌株HB101能够产生最高的质粒单体数量。未来...
由于质粒多聚化是由同源重组引起的,因此缺乏重组酶蛋白 RecA 的细菌菌株的质粒扩增可以降低多聚化频率,并且在某些情况下完全阻止质粒多聚体的形成。DH5α、HB101、NEB Stable、Stbl3等大肠杆菌菌株是recA突变菌株,具有更低的质粒多聚化频率。有研究表明,与DH5α相比,菌株HB101能够产生最高的质粒单体数量。未来需要...
HH-CAS-101p414-TEF1p-Cas9-CYC1t(Plasmid #43802) HH-CAS-100pRPR1_gRNA_handle_RPR1t (Plasmid #49014) HH-ZW-041pYLCRISPR Cas9P35s-N HH-ZW-040pYLsgRNA-OsU6a/LacZ(Addgene 66195) HH-ZW-039pYLsgRNA-OsU3(Plasmid 66193) HH-CAS-130pAc-sgRNA-Cas9(addgene 49330) HH-CAS-129pEM...
图2: 质粒DNA单体(左)与二聚体(右)图谱(图片来源:addgene blog) 1.2 质粒多聚体形成机制:同源重组 1976年,J R Bedbrook等在Cell发表文章:《Recombination between bacterial plasmids leading to the formation of plasmid multimers》,他们发现pMB9质粒DNA在rec+大肠杆菌中呈现出各种不同的大小,更大形式的pMB9...
we integrated the DNA assembly method and the software with a robotic system named Illinois Biological Foundry for Advanced Biomanufatcuring (iBioFAB) (Supplementary Fig.1) to create a nearly fully automated workflow for PlasmidMaker. As a proof of concept, we constructed 101 plasmids across six...
HH-CAS-101p414-TEF1p-Cas9-CYC1t(Plasmid #43802) HH-CAS-100pRPR1_gRNA_handle_RPR1t (Plasmid #49014) HH-ZW-041pYLCRISPR Cas9P35s-N HH-ZW-040pYLsgRNA-OsU6a/LacZ(Addgene 66195) HH-ZW-039pYLsgRNA-OsU3(Plasmid 66193) HH-CAS-130pAc-sgRNA-Cas9(addgene 49330) HH-CAS-129pEM-Cas9HF...
HH-CAS-101p414-TEF1p-Cas9-CYC1t(Plasmid #43802) HH-CAS-100pRPR1_gRNA_handle_RPR1t (Plasmid #49014) HH-ZW-041pYLCRISPR Cas9P35s-N HH-ZW-040pYLsgRNA-OsU6a/LacZ(Addgene 66195) HH-ZW-039pYLsgRNA-OsU3(Plasmid 66193) HH-CAS-130pAc-sgRNA-Cas9(addgene 49330) HH-CAS-129pEM-Cas9HF...
S2, 33 of 101 plasmids suffer either the first or the second situation. More seriously, most nREases are blocked to nick the recognition sites with DNA methylation. In contrast, the nCas9 is not inhibited by DNA methylation as described above. Download: Download high-res image (476KB) ...