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...than AAV vectors. This capacity is large enough to carry the Cas9 gene and a gRNA expression cassette in ...of AdV-modified human cells being subjected to scarless homology-directed genome editing (Maggio et al...

...pU6cm promoter, viral replicon amplification of cargo, and heat treatment.  Figure 1: Left: recombined...and adopts an open conformation in the absence of cargo. bioRxiv 2024.12.20.629623; doi: https://doi.org...

...can be transfected with your transfer plasmid, carrying your gene of interest, and the two helper plasmids...undergoing titering, to remove residual plasmid DNA carried over from the production process. For more details...

... exonuclease, a polymerase, and a DNA ligase to carry out a pooled set of ligations in an hour or less...one to simultaneously complete an ordered set of scarless ligations following digestion with Type II restriction...

...15%, 25%, 40%, and 60% iodixanol solutions are carefully layered and then the viral suspension generated...digested to remove any residual AAV plasmid that was carried over from the AAV production process. Either SYBR...

...of engineered Type IIS restriction enzymes and careful choice of junction sequences guided by experimentally-derived...of overhangs can lead to incorrect assemblies, so care must be taken to minimize this possibility. Recently...

...high-throughput experiments. If a user wants to carry out a high-throughput genetic screen, it is expensive...marker can be removed by Cre-mediated excision for scarless tag insertion.   In addition to the obvious advantages...

...Sharpie to circle 12-48 individual colonies by (carefully) holding plates up to the light, where they appear...diluted from 10X and containing both Hepes and bicarbonate) is brought to room temperature and placed in...

...the Gateway Cloning Platform (Life Technologies, Carlsbad, CA) that represents 180 genes from the Ras signaling... 25877186. 4. Prior, Ian A., Paul D. Lewis, and Carla Mattos. "A comprehensive survey of Ras mutations...

... PITCh plasmic is cotransfected with a plasmid carrying Cas9, the PITCh-gRNA, and the locus-specifc gRNA... Contransfect the PITCh vector with the vector carrying Cas9 and the PITCh- and locus-specific gRNAs Step...

...expect to see for the unknowns in your experiment. Carefully set up a standard curve dilution series that takes...increase the accuracy of your experiment.  Finally, carefully analyze the data and use the trendline that's ...

...numbers for an event you are planning, dangle the carrot (or more popularly- cookies). 3. Do something ...other posts about life at Addgene!Read more of our careers posts!    ...

...translated protein. As depicted in the accompanying cartoon, they have a multitude of uses including (but not...handbooks for protein purification is provided by GE Healthcare Life Sciences. Addgene pages and blog posts ...

...problem-solving process?  You may want to start by carving out time to tackle it. It’s easy to get distracted...been building your creativity all along. In my own career, my team has used these techniques to optimize ...

...Overview An immunoprecipitation reaction is typically carried out in one of two ways. Both methods utilize the...approach appealing, but the disadvantages must be carefully considered before using. For example, the tag ...

... Figure 4: Reporters in Gene On include a mScarlet reporter, luciferase reporter, and neomycin resistance...translation of an array of different reporters including mScarlet-I, luciferase, antibiotic resistance markers, ...

...conferences should strive to provide support to early career scientists, students, and those from historically...build and expand their networks and further their career development, and the scientific community can improve...

... in cold conditions and preferably on dry ice. Careful use of your glycerol stocks When your lab adopts...experiments. The term ‘insert’ refers to plasmids carrying each part, ‘vector’ refers to acceptor plasmids...

...line is maintained for ~20 passages before being discarded and replaced with a new vial of early passage ...produced in the line will be taken offline and discarded. Confirmation of the transfer plasmid Addgene ...

...chromosomal DNA. Chromosomal replication in bacteria is carefully regulated to ensure that it occurs at the appropriate...DNA replication in bacteria, so the process is carefully regulated by a mix of positive and negative regulators...