...PMID: 24901218. PubMed Central PMCID: PMC4047077. 2. Pagliara, Patrizia, et al. "Ostreopsis cf. ovata ...and could be contaminated.Learn how Addgene uses barcodes to track samples Keep track of passage numbers...

...experience and to build upon your existing skills.  2. Find your niche Identify and develop an area of expertise...LLC, a grant professional development firm that includes the social media community GrantChat. She has ... from administration and fundraising to program design and evaluation. She and her team at J. Miller &...

...externally fertilized, transparent progeny (Figure 2). These features give it a leg up (or a fin up?) over...deactivation method in zebrafish are short, modified oligonucleotides called morpholinos. When injected into zygotes...CRISPR/Cas9-mediated insertion of foreign DNA. Despite these caveats, several features of zebrafish—including...

...effective recovery of knock-ins, but did result in 1-2 kilobases of additional sequence being introduced....https://doi.org/10.1093/nar/gku289 Chen C, Fenk LA, de Bono M (2013) Efficient genome editing in Caenorhabditis... DNA double-strand breaks up to 54 bp from the desired insertion site result in efficient editing. Repair...available through Addgene. In 2019, Farboud et al. described another similar co-conversion marker (repair of...whether it be by improved gRNA or repair template design, delivery, screening, and more. Here are some highlights...highlights from the past few years:  Repair template design The Meyer lab meticulously characterized several...of editing. First, they devised guidelines for designing single-stranded repair templates (Farboud et al...

...work. Nobody wins, and you need to make a change. #2 Thou shalt sometimes put work ahead of life Sometimes...that extra two miles which is way more fun. And despite those who may wish to tell you so – towel corners...

... Watch Joanne give her "Not" Networking 101 Talk 2. Tailor every talk to the occasion Don’t just plan...suspense. Notice the simplicity of their slides and how the slides complement the spoken message. Learn from...speak to start creating slides or thinking about content. Create a Google.slides or Powerpoint file right...how these speakers talk without looking at their slides or notes. Listen to their stories and narratives...listeners to be thinking “What happens next?”  Describe how you pushed to a solution or overcame obstacles...term, but are worried that this might sound condescending, consider explaining the term on a slide but...talk will really take and to adjust content and slides to better suit the target time frame.   I suggest...

...of 3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a fluorophore modified from DsRed. DFHO...Aptamer Fluorophores describes the enrichment process used to evaluate oligonucleotides for binding affinity...of Spinach. In doing so, they identified the nucleotides that contribute to the instability of Spinach...similar levels of fluorescence when bound to DFHBI despite being approximately half the length (Warner et ...also smaller than any form of Spinach - only 49 nucleotides in length - and is better able to maintain its... of colors, so do aptamers! Corn was recently described in 2017 (Song et al., 2017). Corn naturally dimerizes... Red and orange fluorescent aptamers Corn was described in the same study as Red Broccoli and Orange Broccoli...

...PubMed PMID: 3459152. PubMed Central PMCID: PMC323600. 2. Chester, Nicholas, and Daniel R. Marshak. "Dimethyl...for the desired modification. Finally, the positive clones are sequenced to confirm the desired modification...Primer Design As a rule of thumb, 11 bp of complementary sequence on either side of the desired mutation...introduced to plasmids using primers (with the desired mutation) in a PCR protocol that amplifies the ...mutated clones. Forward and reverse primers are designed to be complementary, but each primer may extend...Polymerase To ensure that you don’t introduce undesired mutations through the PCR process, you need a ...The resistance marker from the parental plasmid provides a mean for selecting for transformants which have...

...gene1 [lon] ompT gal sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R(zgb-210::Tn10--TetS) General protein expression...and some of the most popular commercial strains designed to avoid them. Why do I need an expression strain...protease: Strains where this is completely deleted (designated lon or Δlon) similary reduce proteolysis of the...which can prevent the culture from reaching the desired density within a reasonable time-frame. For these...and pLysE—the difference being that the latter provides tighter control of basal expression. What if I...don't see protein overexpression? The strains described above should generate sufficient expression levels...you’ve tried a common strain and don’t get the desired level (or any) protein expression? Low expression...

...community is active, engaged, and has great questions! 2. The r/Science subreddit is well moderated. One of...They also host AMA forums where users can ask designated scientists anything about a particular topic....calendar invite over to the r/Science team with a description of our AMA and Joanne’s preferred time to start...was writing all day and straight on through her designated two hours for answering questions. She was able...toxic advisors, fruitless job searches, and even despair at having chosen an academic past. I hope I was...receive via email. The cool features are too many to describe here, but I have over 2,000 notes which Evernote...

... PMID: 9537413. PubMed Central PMCID: PMC3010411. 2. Kohli, Manu, et al. "Facile methods for generating...human cells (Kim et al, 2016). The Waldman lab has designed their AAV tagging vectors (pAAV-SEPT-Acceptor ...vectors for your specific gene, the lab recommends designing homology arms with the modification of interest...combining the AAV and CRISPR-based approaches is described in several recent reviews (Howes and Schofield...

...Nature 339 (1989). 487–488. PubMed PMID: 2725683. 2. Miller, C.J., Kassem H.S., Pepper S.D., Hey Y., Ward...best to start from scratch. Obtain a vial of the desired cell line from a reputable distributor such as ...extreme changes in osmotic pressure and pH can destabilize the viral envelope. One drawback of the multiple...

...AAV variants for widespread gene transfer to the adult brain. Nature Biotechnology, 34(2), 204–209. https...start selecting and testing systemic capsids! Figure 2 contains a decision tree for selecting a capsid, while... highlighted below the table.     Figure 2: This decision tree can help guide your choice on ...expression to specific tissues and cell types. If desired, spatiotemporal control of transgene expression...systemic AAVs to find commonly used vector cargo designs and administration doses. Note: If capsid immunogenicity... Table 1 gives a brief overview of published descriptions of commonly used or emerging capsids. Selected..., B.E., Gradinaru V. Systemic AAV vectors for widespread and targeted gene delivery in rodents. Nature...

...positive cells (Martin et al., 2019). Figure 2: Editing efficiencies using base editing reporters... plasmid delivery to a cell. In contrast, TREE provides a read out for plasmid delivery and subsequent...

...There are approximately 2-4 million proteins per cubic micron in bacteria, yeast, and mammalian cells...with a site mutation (R11G). This site mutation destabilizes the ligase, allowing for active biotin molecules...

... Methods 2009; 6, 343-345. PubMed PMID: 19363495. 2. Wang JW, Wang A, Li K, Wang B, Jin S, Reiser M, Lockey... friends on the Cambridge 2010 iGEM team that describes the basics of Gibson assembly as a parody of “...assembly The Gibson assembly technique was first described by Dr. Daniel Gibson and colleagues at the J. ... Both Snapgene and NEB have tools that help you design primers for PCR amplification of fragments to incorporate...demonstration of how to use Snapgene’s program to design primers for Gibson Assembly. For a simple example...without scars as depicted below. First, you need to design primers to amplify the two fragments while also...the process works best with fragments over 200 nucleotides. This may be because the exonuclease could chew...

... more from the authors in this blog post. Figure 2: iGluSnFR3 displays enhanced brightness, expression...Addgene’s catalog is a new class of sensors for neuropeptides and neurotransmitters. Motivated to develop ...GPCRs recognizing various neurotransmitters, neuropeptides and other ligands - almost A to Z! - from ATP...a new preprint, Jared Toettcher and colleagues describe pYtags, a modular system for monitoring specific... tool for monitoring RTK signaling.  Figure 4: Design of pYtag fluorescent biosensors for monitoring ...iGECInano plasmids here!   Matlashov ME, et al. Design and Initial Characterization of a Small Near-Infrared...intermediate affinities from 40 to 140 mM. These new designs greatly broaden the options for potassium biosensors...

... infected cell and 2. prevent reinfection of those cells. Kaminski et al. designed gRNAs to target the...the virus doesn’t completely disappear; it just hides in latently infected cells. To truly cure HIV-1,... dCas9-SAM to activate HIV-1 transcription and destroy infected cells, the other using wild-type Cas9 ...Why can’t the body mount an immune response to destroy these reservoirs? Essentially, the immune system...enhance transcriptional activation. Zhang et al. designed multiple gRNAs to target the 5’ long terminal ...could be used to combat this problem, just as ART includes multiple drugs to lower the odds of developing...strategy may be a better option than direct cleavage. Despite the potential difficulties in translating these...

..., whereas the mVenus-APT1 fusion does not (figure 2). In the mVenus-APT1, a lipidation motif, which is...any DNA sequence in the way that they like, the design and engineering of fusion proteins is relatively...fusion proteins and highlight some aspects of their design.  Protein size and shape matters The green fluorescent...necessary for post-translational modifications. Despite these potential issues, GFP has been used successfully...consist of random amino acid sequences or (ii) designed to form an inert, unstructured peptide and therefore...C-terminus of the fluorescent protein. I prefer to describe the different parts of the fusion from N- to C-terminus...2011).  Although structural information may guide design, it is advisable to generate multiple constructs...

...as the protocol remains the same.    Figure 2: Microscopy-based antibody experiments should include...controls and be able to use this information to design thoughtfully and thoroughly controlled experiments...is generally a sample or group that will have a desired response. For example, if you are studying the ...sample, the antibody binds specifically to its desired target (purple) and nonspecifically through the...The Preimmune serum control will not bind to the desired target but will bind nonspecifically through the...the Fc, allowing you to detect this undesirable interaction. When non-specific interactions are suspected...nonspecifically with positively charged groups, such as aldehydes in fixed samples. To address this, block positively...

...two visible blue or magenta chromoproteins (Figure 2) (Liljeruhm et al., 2018). This system is a nice confirmation...scarless mutations, ones that don’t leave any undesired scars in the chromosome. These methods roughly... recombination enzymes can also accomplish the desired genomic modification. This technique is frequently...resolution step (“double crossover”) which leads to the desired mutation and excision of the vector backbone. This...the pDS132 MCS with a versatile, synthetic MCS (designed with the help of the DNA tuner program) (Latynski...purified amplicons upstream and downstream of the desired deletion directly into cut vector with excellent... of interest, from the day primers arrive, the desired chromosomal modification can be made in 5 days....

...of concept for further development. Figure 2: Mechanisms of deaminase-based adenine transversion...an abasic site. Like in most other base editor designs, the Cas9 nicks the non-edited strand to encourage...

...become a valuable asset yourself.     Fig 2: A diagram of the Thomas-Kilmann Conflict Model, courtesy...even better. The Thomas Kilmann Conflict Model describes 5 basic approaches to conflict resolution based...

... iRFP713, % Oligomeric state Photo-stabilityc, t1/2, s pKa Brightness in HeLa cells vs. iRFP713, % d ...Biol 24, 758-766 e753 (2017). Baloban, M. et al. Designing brighter near-infrared fluorescent proteins: insights...

...variety of environmental conditions, and double every 2 hours. Yeast are also the first eukaryotic genome ...difficult and time consuming. GAL4/UAS was first described in 1993 by Norbert Perrimon’s lab and has been...

... from damage (Abe & Lim, 2024).    Figure 2: A) When proteins (blue) interact with the air-water... A) Schematic of workflow. B) Detail of spacer peptides (3HB: 11-nm 3-helix bundle; SAH: 60-nm single ...cryo-EM samples from damage at the AWI. Plus, tardigrades!  Find RvLEAMshort here!    Abe, K. M., & Lim...

...ask new and interesting questions in your own lab. 2. Seek multiple mentors during your postdoc At the ...for and how can you do it well? Below, I briefly describe some tips for a successful postdoc fellowship....typically writes the grants that fund your research, guides the research directions of the lab, gives you constructive...Thesis committee members provide expertise in designing and interpreting experiments, as well as career... there a tractable question or are you simply describing a phenomenon? Exploration can be important, but...presentations benefit from practice. If you simply recycle slides from the last time you gave a presentation to a...story I can tell with my data?  Can I make better slides that more effectively tell a story? Generally,...

...experience in cancer research After graduating with a 2:1 BSc in Molecular Biology (roughly a B average in...every item I tested was constantly tracked with barcodes and spreadsheets. It was great to be in the lab...industry positions. There were, however, some downsides to working in a large commercial company. In my...aspects of the company and gave me the freedom to design new projects and grow in the role. I understand... job different every day. I love the freedom to design and develop my own projects, but working remotely...

...Single-chain variable fragment (ScFv) Figure 2: Comparison between the IgG antibody and scFv. ...been infected with a microbe, to trigger immune cascades that clear these infections. Anything that generates...immune response and initiates different immune cascades. In the research setting, antibodies of different...antibody, which recognizes the primary antibody and provides the method of detection. You’ll also want to use...used for. Addgene’s Antibody Plasmid Collection includes plasmids that express antibodies, nanobodies, ...

...Bachelor's degree Generally a short-term position (1-2 years) Often a gap-year position taken to transition... are more involved in the science, experimental design, and execution. Can lead to an independent project...advice Give yourself permission to pursue your own desired path. Careers in science are also not limited to...