...is the most charged, with 5 negatively charged and 2 positively charged amino acids. FLAG tag has been ...(stay tuned for our affinity purification tag blog!).   Applications of epitope tags   The...success rates across all applications. You can find FLAG antibodies on Addgene’s catalog including a mouse monoclonal...protein purification is not ideal with Myc, as low pH conditions are required during the purification, which...experienced. Finding an antibody that works for your application, specifically detects your protein, is species...and have been widely used across species and applications. In this blog, we will review the most common...that are bound by antibodies for the purpose of purification or detection of the tagged protein. These peptides...

...PMC6070312. 2. Stark, Jessica C., et al. "BioBits™ Bright: A fluorescent synthetic biology education kit." ...plasmids and their potential educational uses. We believe that biology education is something that should ... expensive tiny amounts of reagents are, how complicated (and again, expensive) the different machines...desire to create a set of low-cost, easy-to-use educational biology activities – that any school could afford...proteins without the need to handle a living, replicating organism. Furthermore, cell free systems can ...in adapting your research ideas into hands-on educational activities using the BioBits platform, please...BioBits™ Explorer: A modular synthetic biology education kit." Science Advances 4.8 (2018): eaat5105. PubMed...

... remember that a resume is typically no more than 2 pages long. Write your resume for the job posting ...together a grant application? Are you familiar with the publishing or patent application processes? How ... pages in a Curricula Vitae to showcase your publications, academic performances, and research experiences...information. Think "skills," not just "research and publications” What are the skills you developed through your...and writing was how I got my current job as a Communications Coordinator. Wondering how you can develop ...So You Think You have Skills.   Get Science Communication Practice with a Student Group Like Science in...Blogger! Theresa Liao (@theresaliao) is the Communications Coordinator for the Department of Physics &...

... Figure 2: Dense (c), intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, ...infection, because this setup allows for experimental replicates for each of the gRNAs. The idea was to assign...cassette is removed, the effector is expressed. By truncating one of the recombination sites in the construct... reporter (blue) and stained with PI (pink) to indicate cell death. Image from Diessl et al., 2020. ...https://dx.doi.org/10.15698%2Fmic2020.04.713 Truncated TALE-fluorescent protein fusions as a DNA staining...protein to the DNA. They created plasmids for purification of tTALE-eGFP and tTALE-mCherry. In comparison...

...extraction protocol for more details.   Figure 2: An uncut plasmid is expected to form 3 bands: nicked... (AmpR) and origins of replication (ori and f1 ori) which allow it to replicate.   How can I prevent...the bacterial selection marker and origin of replication. During bacterial culture, only the plasmid with...the antibiotic resistance gene and origin will replicate, and the transgene will be lost. Over time, bacteria...the recombined plasmid is still visible. This indicates that the DNA prep contains a mixture of the full...

...MagFRET: The First Genetically Encoded Fluorescent Mg(2+) Sensor. PLoS One. 2013 Dec 2;8(12):e82009. doi: ...Neurobiology has been working on genetically encoded indicators of calcium and other small molecules since the...lymphocytes, too, making them a versatile tool for application in both neuroscience and immunology. Don’t be...imaging for beginners Griesbeck says his calcium indicators can be used in mouse, Drosophila, C. elegans,...give it a shot. “Beginners sometimes think it’s complicated, but actually it is very good,” he said. “It ...his team is now applying it to improve upon an indicator they developed and published in 2010 for visualizing...

...Bacterial Cell Envelope. Cold Spring Harb Perspect Biol. 2(5) a000414 (2010). Cleveland Clinic. Gram Stain. https...ll be talking about the ins and outs of this classification system, including its history, biology, and...repeatable phenomenon, Gram began using it as a classification system. Eventually, any bacteria that retained...permeable to different classes of antibiotics. The classification system is also useful in a laboratory setting...bacteria type. The gram positive and negative classification system is incredibly helpful in visualizing...stepping into a lab.   Anna Kamens is a science communication summer intern at Addgene and a rising third-...

...construct expressing channelrhodopsin 2 (ChR2), a blue light sensitive cation channel, fused to mCherry to the... several months after implantation of fibers, indicating the system was indeed stable for continuous monitoring...interfaces with the entire body, so the obvious next applications are to expand the tool to different brain-organ...

...incorporation of biosensors, protein evolution lineages, 2-photon spectra, more elaborate spectrum analysis tools...non-trivial, however, as FPs are tremendously complicated entities with a large range of characteristics...decision remains spread across literature in publications that introduce these tools. Years ago, Kurt...incorporate FPbase data in their own projects and applications, a REST API allows programmatic retrieval of...

...structure-based framework engineering. Methods, 34(2), 184–199. PubMed PMID: 15312672. Inbar, D., Hochman...antibodies for extracellular and intracellular applications: CDR grafting to stable frameworks and structure-based...and mature HA-tagged proteins in vivo. Nature Communications, 10(1). PubMed PMID: 31270320. PubMed Central...

...peer-mentoring groups at Addgene have gone virtual!   2. Practice public speaking For almost any science career...be a time when you need to speak publicly to communicate your work. One of my 10 tips for giving a great...; Graduate Student Funding Opportunities Open Education Database: 100 Places to Find Funding For Your ...

...yourself greater chances for success.  Build a team of 2 or 3 Don’t go it alone.  You’ll find that no matter...launching a company without partners.  You need a dedicated team that you know well and trust.  This will ...Chowdhury and Clinton Buell, who have very diverse educational backgrounds (an essential component to any successful...

...from a single Illumina MiSeq® run. As shown in Fig. 2, below, differences in input concentration across ...smaller fragments that are then barcoded for identification. This mixture of barcoded fragments is known...of Toronto. He is an author of 20+ scientific publications and patents.   ...

...containing complementary polynucleotide chains. J Virol 9(2): 394-396. Lecomte E, Tournaire B, Cogné B, Dupont...that could be used as a template for downstream applications (Berns, 1972). Using qPCR and restriction enzyme...

... and whether I would be able to carry out a yeast-2-hybrid library screen; instead I thought they would...there a move with either promotions or further education may allow you to transition into various realms...

... 293HEK Inhibits termination step of translation 2-10 ug/mL G418/Geneticin neo HeLa, NIH3T3, CHO, 293HEK... a bacterial origin of replication (ORI) will not allow for plasmid replication in mammalian cells, for... their bacterial counterparts, including how replication occurs and whether selection is necessary for...the nuclear genome, in the absence of plasmid replication, the foreign DNA will be degraded or diluted ...always mission critical to make sure the plasmid replicates in the cell, as many of these experiments are...expressed in trans within the cell for effective replication. Cell lines expressing the Epstein–Barr virus...antigen (293E or 293T cells), allow for episomal amplification of plasmids containing the viral EBV or SV40...

...and in line with Addgene style.        Fig. 2: Reference image (left) provided to our graphic designer...Our educational content  Have you ever wondered how we choose topics for the Addgene blog? Or why we...everything else we do! The goal of Addgene’s educational content Addgene truly believes that sharing speeds...antibody and content team, we identified what applications should be included and what basic information... The guide was then assigned to our science communication specialist, Rachel, to write. Identify expertise...through an editing process and final check before publication.   Do research! Nearly everything we write entails...Guide figures. This requires a fair amount of communication between our content team and our graphic designer...

...cellular physiology.  https://doi.org/10.1002/jcp.30064 2. Barman et al. 2020. “A glance at genome editing with...2020.  “CRISPR Gene Therapy: Applications, Limitations, and Implications for the Future.” Front. Oncol...with a dominant mutation in a gene that causes cataracts could be rescued by coinjection into zygotes of...the technology in different labs for different applications.” Li’s team noted that CRISPR/Cas9 has been ... explore its potential in mice with a dominant cataract disorder caused by a single copy of a gene known...with size limitations. In addition, non-viral applications are also possible through the use of nanoparticles....  “CRISPR Genome Editing Technology and its Application in Genetic Diseases: A Review.” Current pharmaceutical...

...account when simulating a digest as depicted in Lane 2. Controlling methylation Finally, you can control... one half of naturally occurring restriction modification systems that prokaryotes use to protect themselves... has genes encoding one or a few restriction modification systems containing methyltransferases that add... phage for instance). Aside from restriction modification systems, DNA methylation also plays an integral...integral role in regulating genome replication, repairing mismatched basepairs or small indels that occur...methylases) are independent from the restriction modification systems, yet can still affect whether certain...specifically associated with any restriction modification systems, their sequences may overlap with restriction...

...retroviral library was used to transduce T helper type 2 (Th2) cells from mice that constitutively express ..., Nature Communications, 2019. PubMed PMID: 31451697. Genetically encoded voltage indicators for use ... many aspects of ApoB-LP biology, including quantification of abundance, size, and localization in vivo...Pubmed PMID: 23068583 Thierer et al, Nature Communications 2019. Pubmed PMID: 31366908 Adapting sigma54... by Brook Pyhtila Genetically encoded voltage indicators (GEVIs) change fluorescence in real time when... excitation light intensities have made these indicators difficult to work with.  The Gottschalk lab has...