...expressing a selection cassette with 5′ and 3′ flanking sequences matching the sequences upstream and ...to be assayed (e.g. pPD95_86 and pPD57_56), or knocking down YGOI through RNA-mediated interference (e.g... applications used to study gene function. From making transgenic animals, gene targeting and genome modifications...

... used at high concentrations (and vice versa). Looking around the lab, you'll likely find many of the ...mammalian cells.   Name Class Mode of Action*   Working Concentration** Kanamycin aminoglycoside Binds...their mechanism for killing bacteria, and general working concentrations. For instructions on how to prepare...

... no fear - plasmids from Addgene depositors are making it easier than ever to use CRISPR  in E. coli and...gene body can repress transcription by physically blocking the elongation complex from binding the DNA or...Patino R, Heussler GE, Hearne CC, Qu J, Inclan YF, Hawkins JS, Lu CHS, Silvis MR, Harden MM, Osadnik H, Peters... Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim WA (2013) Repurposing CRISPR as an RNA-Guided...

...frequently with little notice or choice Always working on a team/in a matrix - can be frustrating for ...but good performers move around easily  Table 1: Taking it a Step Further - Big Company v. Small Company...on what paths to pursue. Spend some time really thinking about your motivations and what parts of being...

...fewer ethical and logistical challenges than does working with animals. Because Arabidopsis is small, has... to grow, it is an ideal organism for teachers looking to expand their science curriculum. Growing Arabidopsis... biology teacher or professor? Whether you are looking to develop a new teaching resource or you want ...

...replication machinery room to come in and get busy making copies.     Types of origins of replication ...compatibility with one or more other plasmids. Generally speaking, plasmids with the same ori are incompatible because...chloramphenicol – make sure your growth conditions aren't working against you!  The culture inoculum Freshly ...

...Addgene started dealing with this new reality of working remotely, scientists around the world mobilized...took to make all this happen with 80% of the team working remotely. We’re extremely proud of the work we’... maintain our workforce for when we need to be working at full capacity to serve the scientific community...

...from Zhu et al., 2019.  Wensheng Wei's lab at Peking University has developed a new CRISPR screening...injection of precise titrations of AAV or heat-shocking the organism. The SPARC system uses competitive...fluorescing green. Image from Shin et al., 2019. Looking for an alternative to intercalating fluorescent...

...around the world who would otherwise have difficulty making or getting these reagents is profound.” Frank Stegmeier...Library, a library that “enables the high-resolution tracking of more than 1 million cancer cells under drug...became passionate about virology,” he says. He was working in an HIV lab and based on that experience, soon...

...species). This feature is advantageous if you’re working with small capacity viral vectors such as AAV. ... “recovering” its original sequence. If you’re looking to disrupt gene function via indel formation, this...targets. Are you using NgAgo? Let us know how it’s working in the comments section! Read About NgAgo User'...

...In its simplest form, PCR based cloning is about making a copy of a piece of DNA and at the same time adding...restriction site (GAATTC) to the 5’ end of this primer, making our Forward Primer 5'-GAATTCATGTGGCATATCTCGAAGTAC... get PCR amplification. We can start similarly, taking the final 18 bases of the ORF, including the stop...

... Why and Where Do I Find One? If you need help making a transition into a new job or maybe to an entirely...gene delivery, but have limited packaging capacity making them difficult to use with CRISPR as the cargo..... 10. Plasmids 101: Control Plasmids Need help thinking about the types of controls you’ll need for your...

...“reading”) of DNA, have changed our collective thinking about the feasible size and scope of projects ...library prep. As part of our mission, we’ve been working with Addgene to develop and apply our plexWell™...read level, and because custom primers or "primer-walking" reactions are completely unnecessary for sequencing...

...Check out this post for a refresher on AAV titers Making contact with a cell Although an individual AAV ... this type of vector to such a high titer when looking for effective gene expression. How do you achieve... Hanley is a Research Team Lab Tech at Addgene working on methods to improve AAV production.     References...

...hot plasmids articles. Think we missed something smokin'? You can pitch a Hot Plasmids section (antibodies...heatshock promoter) drives expression of the tumor-seeking CAR (chimeric antigen receptor), whereas a ubiquitous... variants with similar capabilities. If you’re looking for a super stable FP for your next microscopy ...

... apart sources of information they don’t like, looking for reasons to dismiss them. There’s evidence to...Zealand’s multiple Covid-19 lockdowns, I held mask-making workshops in my community. I brought along printed... to better meet the needs of the people we are talking to. Emma Harcourt is a PhD student in the Department...

...contributed by Eric Perkins Mammalian genomic DNA is not just a 2-D linear molecule snaking through our cells...These loops not only provide a 3-D structure for packing genomic DNA into nuclei, but are also thought to...

...biological motion (whether endocytosis, vesicle trafficking, or muscle contractions) are produced by orchestrated...the ciliate. It’s thought that they function as a kind of armor to protect their host from predation (Petroni...

...Cas9, and the tracrRNA which acts as a scaffold linking the crRNA to Cas9 and facilitates processing of...different components of CRISPR, It's time to start thinking about how you can used them in your experiments...experiments. Use the following flow chart to get you thinking about the types of experiments you're planning on...

...idea to take some time to confirm that you are working with the correct construct, and verify that the...Consult the manufacturer’s manual for the optimal working conditions for each enzyme. Watch out for methylation...run control uncut DNA to ensure your enzymes are working. When uncut plasmid DNA is isolated and run on ...