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... ask questions like: “Where does my protein of interest live within a cell,” “Does this disease change...Blotting, but it allows you to visualize a target of interest within an intact cell, tissue, or, in some cases...IF, the primary antibody targets the antigen of interest but is not attached to a fluorescent molecule,...products that all claim to target your protein of interest. Thankfully, there are a number of websites compiling...

...is then used to detect the amplified region of interest. Cas13 uses a complementary crRNA sequence to ...based detection of SARS-CoV-2 genetic material (CREST) Sensitivity: 10 copies of RNA per microliter Time...Resources Many thanks to our guest bloggers Shravanti Surest from Iowa State University. Shravanti Suresh is...Sashital Lab, Iowa State University. She is actively interested in science communication and enabling access ...

...selection to isolate plants with the phenotype of interest. Genome engineering, on the other hand, allows...tumefaciens as a vehicle to deliver your gene of interest into a target cell line or organism (Presented...for Golden Gate mediated cloning of your gRNA of interest. Figure 1: Simplified schematic of agrobacterium-mediated...can be used as a vehicle to deliver your gene of interest (GOI) into plant cells. A) The system typically...

...far away brain regions expressing your gene of interest. This can cause issues in tracing studies, where...Promoters The promoter controls how the gene of interest will be expressed; it can be ubiquitous, cell-...particles per volume required to express the gene of interest, in the cells being studied, at an adequate level...Kügler, S. (2017). A MicroRNA124 Target Sequence Restores Astrocyte Specificity of gfaABC1D-Driven Transgene...

...confidently conclude the function of a gene of interest. Tools for RNAi Arguably the greatest advantage...required dsRNA complementary to your gene or genes of interest to trigger RNAi activation. A typical RNAi experiment...that contains sequence against your gene(s) of interest and the reporter. This approach greatly enhances...coupling silencing of APT and any gene target of interest, only organisms that are actively silencing to...

...screens to identify genes involved in a phenotype of interest. This blog post will discuss differences between...target sites occur ~1 in every 8 nucleotides, restricting gRNAs to 5 – 65% of the protein coding region... the lab. Importantly, for any modification of interest, it would be unwise to make conclusions on the...experimental strategy is clear: for any gene of interest, one should require that multiple gRNAs of different...

... other molecule. When paired with a protein of interest, these tools provide an easier means to identify...as empty backbones to clone your own protein of interest. Fluorescence microscopy takes advantage of the...of cellular components, typically a protein of interest. Fluorescent dyes and ligands vary in their chemical...fluorescent ligands can elicit varying degrees of β-arrestin-2 recruitment to receptors, indicating their agonistic...

...comments. Additionally we’ve seen what topics are of interest to you, our readers. Below I’ve compiled a list... from the past year. A quick glance shows that interest in CRISPR continues to grow as the genome editing...community, so email us at [email protected] if you are interested. Addgene's Most Popular Posts of 2014 How...

...N-terminal tags, coding sequences of the gene of interest, C-terminal tags, plant terminators and resistance...component of Golden Gate and GreenGate are type IIS restriction endonucleases. They are like other endonucleases...their own lab: be very careful with the type IIS restriction endonuclease of choice, BsaI, which can be very...

...gene of interest in neurons. This ability to be paired with any Cre-controlled gene of interest makes pmSyn-BFP-Cre...may want to further control where your gene of interest is expressed.  A cell-type specific Cre can provide...

...two phosphatases, PTPN1 and PTPN2, which are of interest as cancer immunotherapy targets but are embryonic...s laboratory at Harvard Medical School and is interested in improving CD8+ T cell responses to cancer....generation of exhausted CD8+ T cell subpopulations and restrains tumor immunity. Nature Immunology 20, 1335–1347...

...expression. Cultures are then lysed and the protein of interest is purified using an affinity tag. Additional ...Determining the best way to express one’s protein of interest can save a lot of time later. Protein purification...C-terminal. Each set of plasmids uses the same restriction enzymes for cloning (see schematic below), enabling...

... to on Twitter? Prior to writing this post, my interest in Twitter was fleeting. I’ve had an account for...Unfortunately, it never clicked for me. Recently my interest was renewed in part due to FOMO  but mostly because... scientist? If you don’t use Twitter, are you interested in getting started? Reply in the comments below...

...With many interesting articles coming out in a myriad of journals every week, it can sometimes be difficult...plasmids and publications relevant to your research interests. We do this by promoting citation, supporting ...encourage you to fill out this form if you’re ever interested in writing your own blog post about a plasmid...

...to determine which job boards list positions interesting to you. Many job postings will result in hundreds...address how you are qualified and why you are interested specifically in the position. We frequently receive...cover letters. It is also important to express interest, even excitement, in a position. Everyone prefers...

...somewhat unusual things. For example, I am not that interested in meeting movie stars, but when I met the astronaut...a scientist up there, but the panelists were interesting enough to make up for the somewhat naïve questions...their seminal, cooperative paper in Science.  Interestingly, this project started with their labs studying...

...NEBCutter2: For finding restriction enzyme cut sites Webcutter 2.0: For finding restriction enzyme cut sites ... different types of cloning methods, such as restriction enzyme cloning, Gibson assembly, or In-Fusion...

... Short segment of DNA which contains several restriction sites allowing for the easy insertion of DNA....“vectors” or “constructs.” To insert a gene of interest into a vector, scientists may utilize one of a... a variety of cloning methods (restriction enzyme, ligation independent, Gateway, Gibson, etc). The cloning...

...figures of interest along with the article metadata, legend and user created notes on a “Pinterest for scientists... scientific literature? Very often the data of interest are in 1 or 2 figures from one paper and then ...

...collection, you'll likely find some proteins of interest for which you have several antibodies in stock...another, or if you’re looking for a protein of interest while it’s in a transition state or specific configuration... to validate that it recognizes the protein of interest in specific configurations or conditions. In those...

...easiest and clearest ways to determine if your sample has recombined. Choose a restriction enzyme that...one resulting plasmid carries the transgene of interest (aka the viral genome) and the other contains ...

...2008 the Quake Lab at Stanford University became interested in exploring biological dark matter – large tracts...like YNP thermal sites - are also of practical interest since they’ve previously yielded valuable enzymes...bacterial RNA-Seq. This enriches for transcripts of interest without an increase in the number of steps in ...

...overcome. Conflict, and its inherent tension, is interesting. It’s what drives a story forward - and this ... the pieces that make you think, “Oh, that’s interesting! How did you solve that problem?” Those are the...your audience in with something that sparks their interest and makes them want to know more. This could be...

...plasmid that produces biotin ligase and a protein of interest fused to maltose binding protein (MBP, New England...-Avi-His/BirA to add biotin to your protein of interest Grow fusion protein plasmid in NEB T7 SHuffle... Vanderbilt Eye Institute and is particularly interested in cell signaling.     Additional Resources on...

...several excellent guest posts from our depositors. Interested in writing your own post for the Addgene Blog...(AAVs) are commonly used to deliver a genes of interest to a variety of cell types in-vivo. These are ...customizable functions to your DNA sequence of interest. Potential applications of the system include ...

...clone 500-700 bp homology arms for your gene of interest into the FP-SEC vector using Gibson assembly. ...as well as a promoter-FP fusion for the gene of interest. L1/L2 larvae from these strains are then heat...applications, SEC selection could replace other, more restrictive C. elegans selectable marker systems. This novel...

...identified some fly stocks and other reagents of interest, the next question to ask is what to do with them...fly line expressing Gal4 in your tissue type of interest. You can quickly mate your UAS-YFG line with a...different tissue types! Although putting a gene of interest directly downstream of a tissue-specific promoter...

...all viral genes are flanked by EcoRI and BamHI restriction sites - so cloning projects should be fun and...researchers. We have been excited to see so much interest, because it means more scientists will be working...Communications Team at QBI UCSF. They are particularly interested in facilitating collaborative science, and making...

...determine if the fluorescent proteins you are interested in are compatible with your microscope optics...related protein, if you know that your protein of interest has a similar spectrum. For example, here's a ...need to try several proteins in your system of interest. Finally, in this set of proteins the green and...

...first developed in the 1990s. While traditional restriction enzyme cloning uses short sticky ends to insert.... To start the SLIC cloning process, a gene of interest is PCR amplified to add 5′ and 3′ homology regions...multiple fragments without the need for specific restriction sites or DNA ligase! This post was originally...

...efficiency. However, gamma-retrovirus infection is restricted to dividing cells only, so is not suitable for...can target the virus to specific cell types of interest. This is a particularly nice feature as organ-...have an idea which ones could be helpful to you. Interested in learning more about a specific virus? We have...

...tumorigenesis The conference opened with a very interesting presentation by Professor Guido Kroemer from ...presentation, starvation or the use of caloric restriction mimetics trigger autophagy and, in a mouse model...References: 1. Pietrocola, Federico, et al. "Caloric restriction mimetics enhance anticancer immunosurveillance...

... indels for “functional knockout” of a gene of interest is one of the most common applications of precision...important difference (Figure 3). If you are interested in harnessing the power of digital PCR screening...Postdoctoral Fellow at the University of Alberta. He is interested in everything molecular biology. Follow him on...

...activated B cell that produces the antibody of interest with a myeloma cell. This RNA was then used to...Constructs were then verified by both diagnostic restriction digest and colony PCR.   Fig. 1: (A) ... found that the recognition sequence for the restriction enzyme BciVI was present in the aberrant light...

...the bacteria can easily uptake your plasmid of interest. Scientists can also use electroporation, the ...scientists to introduce different DNA sequences of interest into a bacterial cell or a host’s genome. To do...create both transient cell lines, where a gene of interest is expressed but not integrated into the genome...

...coli, carrying a plasmid expressing a protein of interest). Mesophilic microbes are highly preferable (and...delighted to engage with potential collaborators and interested audiences. Please visit our team wiki or contact...at the iGEM Competition 2019. He has an active interest in scientific writing and science communication...

...been hampered by the need to tag each protein of interest and, for in vivo work, generate a dedicated transgenic...GFP-nanobody rather than a specific protein of interest. This fusion protein can then be co-expressed ...expressing a GFP-tagged version of a protein of interest. The binding of the GFP nanobody to its target...

...Researchers express genes of interest from plasmids in order to study gene function or to engineer cells...fluctuations due to copy number by introducing genes of interest into the chromosome. But this has its own set ...thereby intensifying repression of the gene of interest and keeping its expression level constant. In ...

...Health Science Center at Houston. Prakash became interested in studying TS when he was studying thoracic ... Dr. Carolyn Bondy, and Dr. Angela Lin got me interested in TS when I was a newly independent faculty ...channels in the brain.  “I have a long standing interest in the biology of GPCRs and G proteins, with an...

...doing after the injunction? We are particularly interested in whether institutions planning to raise salaries...our FOIA requests, we are starting to see very interesting data on how much postdocs are actually getting...Executive Director of Future of Research. He is interested in advocating for and with junior scientists,...

...fail. Our laboratory manager would definitely be interested to see which part of her procedures are the most...getting up and running and more time working on interesting biotechnology code.There are valid technical ...Shamloo is a Software Engineer at Addgene and is interested in writing software tools to help scientists....

...them a quick scan for interesting key words and download what seems interesting or relevant. Harness the...keeping up with the literature in your field of interest. Set aside some time each day or week to at least...

...When this interaction occurs, BirA* activity is restored, and nearby proteins are labeled with biotin. ...difficult to detect proteins with short half-lives. Interested in using Split-BioID with your research? Plasmids...

...which may be of particular interest to MoClo users!). If you’re interested in complex Golden Gate Assembly...overhang set by uploading the set with the Type IIS restriction enzyme and thermocycler protocol. It will also...

...in plants to conditionally deplete proteins of interest in C. elegans (Zhang et al., 2015). In this system...spatially and temporally knock-down proteins of interest.  Plasmids for cGAL and split cGAL systems The...halves of cGAL can drive expression of your gene of interest.  SapTrap Tool Kit for CRISPR Cas9 edits At Addgene...

...potentially signaling a rough road ahead for students interested in leading the process despite the recent advance...students will pursue this newly available avenue. Interestingly, both of these changes share a common thread...understand human disease.  Outside the lab, he is interested in communicating science and in the intersection...

...integrity of your virus, perform a diagnostic restriction enzyme digest and compare it to some reliable... use the virus immediately upon harvesting. Interestingly, freeze-thawing host cells immediately prior...24003179. PubMed Central PMCID: PMC3760253. 2. Rohwer, Forest. "Global phage diversity." Cell 113.2 (2003): 141...

...and type of target sites that flox your genes of interest, a powerful FLEx switch can be created for your... considered "Cre-On" meaning that your gene of interest starts in the inverse/antisense "off" position...can be employed for "Cre-off" (i.e. your gene of interest starts in the sense orientation and Cre is used...

...chance as entrepreneurs. To make things more interesting, we are working in a new technical discipline...things we could do for work, and they weren’t restricted to science jobs. Keira’s an optimist, I’m a realist...doing makes sense. I can support that.” That was interesting. If people could be against GMOs as food but ...

...gradients use dilutions of your sample or protein of interest to understand the dynamic or working range of ...particularly in the sizes around your protein of interest, but you don’t want to run so long your dye front...membrane. If you’re not sure if your protein of interest is over-transferring (going completely through... validated for your protein and application of interest or validating an antibody yourself. But even with...