... recruit your protein of interest, maybe you want to inhibit it. Restricting the effect to a specific ...cell morphology and adhesion But what if you’re interested in that other most-important component of the...knockout is not feasible or when the protein of interest supports a variety of processes in different cell...clustering. Fuse either CRY2 or CIB1 to your protein of interest and express the other separately, then use blue...where toxicity doesn’t matter. Plus, for folks interested in directed-evolution methods, the activity-based...

...recognize on a host cell. To make matters even more interesting, much of what a phage faces as it approaches ...abilities (6). This can be accomplished using a restriction enzyme with a short recognition sequence (like...with incubating this membrane with your cells of interest and growing them up overnight. Assuming they expressed...that their host colonies grew. If your cells of interest stick to the same location as a given protein,... located. Now you can grow up your colonies of interest, sequence the inserted DNA and characterize your...Use the FX Cloning Sytem to Clone Your Gene of Interest Into a Variety of Expression Vectors Deposit Your...

... spectrophotometer Restriction Digests Cut and analyze plasmid DNA with restriction enzymes Agarose Gel...pooled-plasmid libraries Diagnostic Restriction Digest Verify a plasmid using restriction enzymes Watch the Video!...new window) Link to Video Restriction Cloning Assemble plasmids by restriction digest and ligation Cloning...Cloning by PCR Generate restriction sites by PCR Modification by Annealed Oligo Cloning Add a short stretch... Cloning (LIC) Scarless cloning with Type II restriction enzymes and T4 polymerase pLKO.1 - TRC Cloning...

...1. Getting your viral vector into your cell of interest: AAV tropism and route of administration AAV tropism...Table 1 from the paper. 2. Manipulate your cell of interest: Cell-type specific expression of molecular tools...approaches to control transgene expression by restricting expression to genetically defined cell types....can then be targeted with optical stimuli. The restricted light exposure to a specific area in combination...brain area can then reveal the cells bodies of interest. The toolbox for neuroscience is becoming more...

... gene of interest to either the N-or the C-terminus. Even if a researcher’s gene of interest isn’t in ...in the same vector backbone with very similar restriction enzymes. You can essentially cut and paste quite...mind that a very simple sequential or double restriction enzyme digest will open up possibilities exponentially...here). If you’re not sure where your protein of interest localizes in the cell, use these markers as your...

...with certain scientific areas. Whether you’re interested in “big P” or “little P” policy, the possibilities...career pivot. There are many ways to pursue your interests in policy or advocacy as a supplement to your ...are many ways to make it work for your time and interests.  Action Items:  Sign up to receive updates from... 2). Starting early demonstrates not only your interest to future positions, but will also make your transition...someone working in a science policy space you’re interested in. Many fellowship programs can connect you ...health services research. She is particularly interested in driving scientific and technological innovation...

...you’re interested in (i.e. biotechnology, pharmaceuticals, etc.) Do their jobs sound interesting? Is their... if a master’s might be for you. 1. Are you interested in master’s-level jobs? Ultimately a degree is...website like Indeed. Do the job descriptions sound interesting? Is the work something you can see yourself enjoying...figure out if a master’s is a good fit with your interests and career goals. You might still decide to earn...

...guest blog pieces. Throughout my internship, my interest in scicomm has grown and now it feels like an ...away-from-the-bench career option. So if you’re interested in learning more about science communication ...I just keep learning on the job. It keeps it interesting.” And while his official title may be science...nice is a great way to bring people in.” Are you interested in careers outside academia like science communication...If so, let us know what types of jobs you are interested in in the comments. They could be featured in...

...who are interested in understanding the work you do, in addition to being genuinely interested in knowing... asked about my current projects and personal interests. We chatted about social media analytics and summer... There was something so special about her pure interest and curiosity to understand my work and me, especially...to-dos, I also found time to go out to sushi restaurants for lunch with coworkers and visit the office...

... inverted repeats (green) flanking the gene of interest (blue) in a plasmid backbone. A separate plasmid...the TA sequence is duplicated near the gene of interest insertion site.   The Sleeping Beauty transposon...RFP, no FP). The vectors each contain two SfiI restriction sites with different overhangs depending upon...whether they are on the 3’ or 5’ end of the gene of interest allowing for insertion in the correct orientation... lines which can robustly express your gene-of-interest, either constitutively or regulated by doxycycline...

...matters! There are important societal reasons, self-interested reasons, and, given that science is largely funded...scientists to communicate about their work and their interest in science more generally to people outside of...scientist, start. If they don’t seem all that interested in the details, work on telling it another way...such as COMPASS. Although COMPASS has a special interest in ocean science, they have good advice for anyone...blogging here, just leave a comment if you're interested and our team will get back to you.  ...

...impactful research topic that aligns with your interests. We've outlined some important factors to consider...brand. It offers colleagues insight into your interests and will determine how you spend a good chuck ...career by connecting you with people of similar interests and goals. The following questions can serve as...journals that discuss general science in the area of interest. “Have a look at what others have done recently... your project will be fresh as well.” Don't underestimate the importance of choosing a solid research ...

... such as Stbl3s. ITR loss can be detected by restriction digest using enzymes, like SmaI, which have recognition... such as SaCas9, CRISPR gRNAs, or your gene of interest. Promoter The promoter drives the expression of...organism or tissue type-specific, which can help you restrict the expression of your transgene to a specific...result is a permanently flipped transgene. By restricting the availability of Cre, lox sites lets researchers...successful and specific expression of your gene of interest. References Haery L, Deverman BE, Matho KS, Cetin...

...of knowledge or interest in STEM subjects; we generally refer to them as the “interested public.” Subscribers...to Scientific America would be considered the interested public and it would be safe to assume that they...age range, familiarity with the subject, shared interests, socioeconomic and/or racial/ethnic background...example adults 45-55 yrs old who breed or are interested in breeding dogs who also enjoy listening to ...

...need to add unique restriction sites on the ends) and amplify your sequence of interest with Taq polymerase...cloning) is a DNA cloning method that does not use restriction enzymes or ligase, and requires no post-PCR procedures...depicted as a green cloud) functions both as a restriction endonuclease and as a ligase by cleaving and ... confirm the presence of your insert by PCR, restriction digest, or sequencing. Pro tips Do not add 5...

...one problem: how to get that into the cells of interest,” Griesbeck said. Because they are chemical compounds... to go. Griesbeck is motivated by a particular interest in monitoring the activity and biochemistry of... Technische Universiteit Eindhoven has similar interest in FRET-based sensor proteins, particularly for... doi: 10.1371/journal.pone.0082009. (PubMed) Interested in More Hot Plasmids from Addgene? Check out...newly available plasmids at Addgene. If you're interested in plasmids for in vivo imaging, check out this...

...antibodies could thus be interesting drugs. Known drug targets  Another interesting approach is to design...expression dataset are causal for a phenotype of interest. Selected pathways  I attended a great talk when...to calculate which would be the best gRNAs. Interestingly, the libraries do not overlap much, showing ...identify all the alternative names for your gene of interest (that were not found by Green Listed), and test...Karolinska Institutet, in Sweden. He has a particular interest in using molecular tools to study the immune system...

...interactions is the pull-down assay, wherein a protein of interest can be captured along with its associated partners...chromatin fragments that are bound by a protein of interest are pulled-down and later coupled to high-throughput... the antibody problem is to tag the protein of interest with a small biochemical “epitope tag” such that...generated, each with a different tagged protein of interest, and a universal all-purpose “tag antibody” can...crRNA (guide RNA sequence targeting the gene of interest) and ssODN (tag-encoding donor DNA) using our ...Regenerative Medicine (CRM), Edinburgh. His research interests include optimizing CRISPR/Cas9 genome-editing ...

... often struggle to understand how a protein of interest actually behaves. How large is it? What parts ...required is that the proteins/nucleic acids/lipids of interest are fluorophore labeled and that these fluorophores...each other. How do you label your biomolecule of interest with FRET fluorophores? When is absolute labeling... FRET acceptor-binding site on your protein of interest: you just don’t know it yet! The poly-histidine... Addgene, where you can insert your protein of interest, resulting in a C-terminal fusion of a tetracysteine...Brigham and Women’s Hospital in Boston, MA. His interests include developing novel fluorescent tools for...

...tool that allow scientists to express a gene of interest in a cell or animal model. AAV plasmids contain...cassette, which contains a promoter, the gene of interest, and a polyA signal. Created with biorender.com... an AAV vector plasmid, they flank the gene of interest. A single ITR consists of palindromic arms (A-...identification of non-dominant ITR sequences.  Restriction digests with the enzymes SmaI and SrfI can also...deletions and point mutations will go undetected by restriction digest methods.  This blog post was written ...