... are nuclear- or DNA-binding proteins, you may need to sonicate your lysates in order to release the proteins...ladder on your gel and keep an eye on it and the leading line (dye front) visible in the sample buffer. ...stripping and reprobing. Just make sure to go from least to most abundant protein if you’re stripping, since...

...fantastic model organism for many reasons, not the least of which is ease of transformation. There are many...finding true transformants more difficult. Don’t leave your plants on selective plates for too long: even...recovered from the stress of selection you can clip a leaf and extract genomic DNA for genotyping. This won...

...commonly result in a complete loss of function while nuclease-deactivated Cas9 (dCas9) can be tethered to a ...which contains multiple point mutations in the nuclease domain. These mutations result in an inactive ...about CRISPR Kinome Libraries available at Addgene Learn about the advantages of pooled CRISPR libraries ...

... in the primary literature, and as a result can leave scientists under-informed with respect to how different...the year, those that meet our minimum standards (clear and detailed methods and image descriptions, a minimum...computer files. If you asked your team members or colleagues: How much unpublished AAV data do you have sitting...

...retroviruses. Lentiviruses are able to enter an intact nuclear membrane allowing them to infect both dividing ...Third-generation plasmids can be packaged with either system. Please note that the generation used to package the virus...Addgene Blog: Download our eBook: Plasmids 101 Learn about genome-wide screening with CRISPR and Lentivirus...

...this gap by both highlighting that researchers do learn MANY transferable skills while working in the lab...to live in your lab, right? Wrong! While it is clearly a good idea to be diligent in your research, remember...career center. Alternatively, it might even be a colleague. A mentor does not necessarily need to be someone...

...subsequently changed fields and my cohort of friends and colleagues had moved on as well, so I knew very few people...people, it was certainly an issue worth addressing. Clear expectations have been established for future participants... after our initial pilot, and look to take some leaps. Binning Singletons is its own organization, working...

...CRISPR 101 Series Learn about the PAM Requirements of Different Cas9 variants Learn about CRISPR Pooled.... & Liu, D. R. Genome editing with CRISPR–Cas nucleases, base editors, transposases and prime editors....

...much trial and error to learn the scripting language, but our path forward was clear: to build one process...assembly algorithms meant that we already had a clear idea of the necessary steps to perform:    ...

...been demonstrated in zebrafish using zinc-finger nucleases, TALENs, and more recently CRISPR/Cas9. Another...zebrafish zygote transforms into an organism with a clearly established body plan including head, eyes, and... tracked in whole brains of swimming, actively learning larvae, providing unprecedented views into cognitive...

... variety of expression systems. Keep reading to learn more about the Gateway cloning method and its advantages...conditions, the attL and attR sites can recombine, leading to the excision of the phage from the bacterial...Resources on the Addgene Blog Try Out FastCloning Learn About Methylation and Restriction Enzymes Use Colony...

...contributed by the Mesothelioma Cancer Alliance. We have learned much about the causes of cancer and the different...nucleus, if the cell dies from injury, this protein leaks out into the cytoplasm, migrates out of the cell...About Cancer and Addgene's Cancer Collection Pages Learn How the ClonTracer Library Can be Used to Study ...

...for Titering Your Lentivirus Learn All About Viral Vector Elements Learn How to Do Genome-Wide Screens... target cells. Poor tissue culture practices can lead to low titers and poor transductions. Cell lines...

..., using up valuable resources in the cell thus leading to slowed growth. Additionally, some protein products...well for most experiments, the lac promoter can “leak,” meaning that a low level of expression exists ...your favorite protein expression tips and tricks! Leave a comment below or browse Addgene’s collection of...

...a first time AAV user Learn about how AAV is used in retinal gene therapy Learn about the parts of an ..., would help improve split vector strategies and lead to greater and more consistent transgene expression...

...very high titers (Zingg et al., 2017), which can lead to cells in far away brain regions expressing your...experiment. Some inducible constructs can exhibit leaky expression in the absence of the inducing drug, ... result in overexpression of your gene and could lead to toxicity or trouble interpreting your results...

...temperature 5-10 degrees above the primer and be at least 25 bp away from each primer, ideally located close...suitable for your cell lines and/or tissue types.  Please do note my oh-so-subtle use of reference genes,...can be used up quickly by abundant transcripts, leading to falsely lowered levels for less abundant transcripts...

...episode of our Hot Plasmids podcast series, you'll learn about optogenetics tools for controlling protein...plasmids covered in this segment Hot Plasmid 1: Photocleaveable PhoCl protein for controlling molecular processes...

...me: Drag-and-drop genome insertion, without DNA cleavage, with CRISPR-directed integrases🎵 On the 6th ...Addgene gave to me: dCas9-based gene editing for cleavage-free genomic knock-in of long sequences🎵On the...

...Guide to Synthetic Biology One of the best ways to learn about a new field is simply to know where to look...have additional resources that might be helpful, please reach out to me by email.  About the author Niko...