...created with the same backbone but that contain different inserts. When you design primers to anneal outside...primer pair to screen for the presence of many different inserts. The downside: this type of primer does... The type of primers you use depends on your preferences. Either way, make sure to test your colony PCR...a standard PCR thermocycling program. One key difference is the plasmid DNA must be released from the ...and without an insert. These controls are quick references you can use when you run your PCR products out... an appropriate molecular weight standard for reference and to add a loading dye with glycerol to your...products have any mutations.  There are lots of different cloning strategies, but regardless of which is...

...the integrity of different targets and labels. You may want to check out a few different protocols to look...you’ve been looking for too!     References and resources References Bertiaux E, Balestra AC, Bournonville... two points within ~250 nm of each other. For reference, individual presynaptic terminals, which are jam...optically magnify quite so much. Created with biorender.com Steps 2-4 are not found in standard IHC/ICC...traditional full IgG secondaries, can improve signal strength (Sherry and Stiles, 2022). Fourth, signal continuity...2020). Plus, it’s been demonstrated to work in different model systems, paired with SRM techniques, and...

...be translated to other lines. Differences in the physiology of different cell types, however, can make...not only between serum from different manufacturers but also between different lots of serum from the same...concentrated dilutions you will be able to overcome differences in receptor expression. It's all in the mix Even...typically employ the positively charged polycation, polybrene. Interestingly, one study found that alternative...Virus? Check out Addgene's New Viral Service! References 1. Hay, R.J., Macy, M.L. & Chen, T.R. “Mycoplasma...tropism of lentiviral vectors through pseudotyping.” Current Gene Therapy 5.4 (2005). 387-98. Pubmed PMID: 16101513...

..., many plasmids were available, but they often weren’t easily accessible, due to a wide range of logistical...world. This idea, that available and accessible weren’t always the same thing, shaped both Addgene’s founding...distributor for that country. In China, direct shipments weren’t able to clear import control. Partnering with a...form of problem-solving. When universal solutions aren’t enough to ensure accessibility, partnering with...particular topic into beautifully designed and easy-to-reference eBooks. Figure 3: Global readership of the Addgene...which received over two million views from 119 different countries (Figure 4). Four of the five most popular...researchers who use them. The best part of any conference is scientists, from techs to grad students to...

...shared ownership. Go check it out! References and Resources References Radtke, A. J., Chu, C. J., Yaniv,...mean highly multiplexed: you can image over 65 different parameters using this method (see Movie S8 from...for spatial biology enthusiasts. This dataset currently includes 1049 community-validated reagents from...fluorescent probes, representing imaging from 47 different tissues. Researchers upload and share both their...resources .csv file for easy filtering and sorting. Reference images can be viewed by clicking on the links ...data, more discussion, and more functionality. Transparency, both in what works and in what doesn’t, and...

...works well to generate libraries, they all have inherent shortcomings compared to one pot saturation mutagenesis... and scalability (see Supplementary Table 1 of Wrenbeck et al). It uses a uracil labeled wild-type ssDNA... saturation mutagenesis technique described in Wrenbeck et al is a PCR-based approach for generating a...264 basepairs, was the maximum length tested by Wrenbeck et al but it’s likely that longer stretches of...of positive clones. See Supplementary Note 1 in Wrenbeck et al for details on how nicking was optimized...mutagenesis? How would you use it in your research? References 1. Natarajan, Chandrasekhar, et al. "Epistasis...Protocols, pg. 103-109. PubMed PMID: 20676978. 3. Wrenbeck, Emily E., Justin R. Klesmith, James A. Stapleton...

...construct containing fluorescent proteins (XFP) of different colors to label an organ or entire organism. Because...localization sequence, allowing it to localize to a different area of the cell than CFP and YFP. The most recent...could recombine to activate the expression of a different fluorescent protein. Therefore, each cell could...Confetti mouse was first used to describe the self-renewal of cells in the intestinal crypt, and since then...proliferating cells. Tetbow optimizes Brainbow for current technologies Tetbow (Sakaguchi et al., 2018) overcame...method that makes tissues, such as the brain, transparent so that the 3D architecture of a fluorescent ...can be combined with new imaging techniques.  References Cai, Dawen, et al. "Improved tools for the Brainbow...

...however different proteins will likely achieve different levels of incorporation due to differences in stability... further technical reference and protocols for use of this system, please reference ‘Rinehart Lab reagents...tools such as phosphospecific antibodies, which aren’t available for the majority of documented phosphorylation... length of induction, which will vary between different proteins. Beyond these standard considerations...mitochondrion. You can follow her on Twitter @nieminm.   References 1. Cohen, Philip. "The origins of protein phosphorylation...

...of interest supports a variety of processes in different cell types. One great way to do this is to use...model for optogenetic studies since they are transparent and the developing embryo is accessible for injection...SuperPLD module is optogenetically recruited by different CIB constructs to the plasma membrane or lysosomes...future updates to the collection! Resources and references Additional resources on the Addgene blog Viral....org Optogenetics Guide Optogenetics Plasmids References Aljiboury, A. A., Ingram, E., Krishnan, N., Ononiwu...precise control over vascular endothelial barrier strength. eLife, 12, RP84364. https://doi.org/10.7554/eLife...optogenetic tool for localized, microtubule disassembly. Current Biology, 32(21), 4660-4674.e6. https://doi.org/...

...hard to count the number of different commercial strains of E. coli currently available – a quick Google...the intestinal tract of animals. There are many different naturally occurring strains of E. coli, some of...more convoluted due to researchers sharing and renaming samples throughout history. It was likely isolated...tetracycline Additionally, Table 2 provides a quick reference for some of the popular strains, their genotypes...lacZΔM15] hsdR17(rK-mK+)  MC1061 Streptomycin Parent of DH10B/TOP10 and derived strains, common lab ... has a great list of genetic markers for your reference. Browse Addgene's curated list of Bacterial Expression... Matthew Ferenc...

... to use pre-drawn images in your figures Biorender Biorender is like clip art for scientists. The web-...guarantee Biorender will create it. Additionally, you can upload your own images to the Biorender workspace...workspace. Free Biorender accounts can store 5 workspaces at a time and workspaces are only downloadable in...in a small file size with a Biorender watermark, which is usually ok for informal settings like lab meeting... paid account.  I’m proud to announce that @BioRender (a project my team & I have poured our hearts ...

...subclass, and are part of the reason why different isotypes drive different types of immune responses. Handily... do so much more than that!    References and resources References Clynes RA, Towers TL, Presta LG, ...immune cells. (psst! Need a quick review of the different immune responses? We got you! You can also review...receptors; and so on and so forth. There are a lot of different receptors, but their downstream effects can be...region contributes to affinity and specificity. Trends in Immunology, 29(2), 91–97. https://doi.org/10.1016...

...Laureates, who speak about their work and current, trending topics in their area of research. The society...students with their applications. The society currently has 1,283 members, who comprise undergraduates...students who want to study chemistry. The society currently has 352 members, ranging from undergrad all the...Biological Society based at the University of Oxford currently has around 700 members and hosts a range of scientific...provides talks and career support for a range of different scientific disciplines, as well as social and ...members as well as arranging collaborations and conferences for their students. Another large goal of the...

...something you CAN choose and it will make a huge difference for your future career and life. You might see...amazing scientists, but the labs were profoundly different. My first rotation was with a mid-sized lab with...hone in on the factors that will make the most difference for your success. Gather data on the lab choices...? What is the energy level in the lab? Does a frenetic atmosphere make you edgy or energized?   Are you...used to a lot of attention? Or are you one of 8 children and used to getting the attention you need? Are...your best performance and learning? Everyone’s different so give some thought to what atmosphere makes ...lifelong learners. I believe we won’t be happy if we aren’t learning so before you take any position you must...

...plasmid sample has a mix of different-sized multimers, you may even see different sets of bands on the gel... us at [email protected]. Resources and References References Bacolla A, Wang G, Jain A, Chuzhanova NA...multimerization has been observed to occur in many different kinds of plasmids, taking many forms (Levene, ...from BiteSize Bio for more information on how different plasmid conformations run on an agarose gel). ... tips. Portions of this figure were made with BioRender.   Grow plasmids in recombinase-deficient...Plasmids 101 eBookAddgene's Molecular Biology ReferenceAddgene's Restriction Digest Protocol More resources...

...view previous versions.  Currently, all submissions also require a DOI referencing primary literature, and...the time) began an effort to collate some of the current “best” FPs, and I collaborated with him on a web-based...that one might like to store about FPs than is currently represented at fpvis (complete spectral information...sequences and lineages, FRET efficiencies, additional references, etc...) and a more robust database structure...address).  One may also simply attach important references that serve to further characterize a protein ...plenty of room for improvement and elaboration.  Current projects include incorporation of biosensors, protein...

...cells (Ma et al., 2015). To do so, they fused a different fluorescent protein to various Cas9 variants, ...either MS2, PP7, or boxB) that each recruits a different fluorescent protein (BFP, GFP, or RFP) to the ...green, or red readout. But gRNAs containing two different hairpins will produce a secondary color (yellow...allows for simultaneous visualization of sites on different chromosomes in living cells, the system requires...they chose an intergenic DNA region (IDR3) as a reference loci to measure inter-locus distances of loci ...sgRNA-Sirius-4X(MS2-PP7) showing recruitment of different fluorescent proteins. (B) CRISPR-Sirius used to...try. Find CRISPR-Sirius plasmids at Addgene. References Ma, Hanhui et al. “Multicolor CRISPR labeling...

...efficacy of different materials so that we could have discussions about the pros and cons of different fabrics...about the difference between airborne and aerosolised droplet transmission and how different fabrics and...sexual and reproductive health.    References and Resources References Drummond, C., & Fischhoff, B. (2017...used to stay up past my bedtime, poring over my grandparents’ medical textbooks by torchlight under the covers...

...in 1983. The differences between gram positive and negative bacteria is due to differences in cell wall...minoring in studio arts photography.  References and resources References Silhavy, TJ., Kahne, D. & Walker ...negative and positive bacteria are permeable to different classes of antibiotics. The classification system...system is incredibly helpful in visualizing differentiations between bacteria, leading to safer medicine...diagnostics/22612-gram-stain Hardy, J. Gram’s Serendipitous Stain. https://hardydiagnostics.com/wp-content...

...emission wavelengths. Renilla luciferase from the sea pansy (Renilla reniformis) emits a blue light with...(Vargulin) luciferin None Yes Sea pansy(Renilla reniformis) 480 36 Coelenterazine None No Deep sea...laser to rapidly excite GFP. As a result of the different mechanisms to produce photons, chemiluminescent...Cofactors/Substrates Required Not Required Signal Strength Lower Higher Sensitivity Higher Lower Background...