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  1. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

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    Blog Post
    ...rate as compared to a constitutive promoter. To simulate the increase in plasmid copy number during the...expression from the constitutive promoter did not. Similar levels of expression between a plasmid and the ...stabilized promoter, deoxychromoviridans titers were similar regardless of where the genes were expressed. What...
  2. Mouse Modeling, Part 1: Genetically Engineered Mice

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    Blog Post
    ... a way to track successfully engineered cells. Similar to transgenic mice, knockout mice are engineered...off-target effects on regions of the genome with similar homology to the site of interest or create mosaic...recombination of floxed sites in the cell can occur. Similar systems include tetracycline or doxycycline mediated...
  3. Making CRISPR Plasmids Using Fragmid

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    Blog Post
    ... components. Using the handy website, users can simply select the components they want for each module...license.   Once you have the components you want, simply assemble them into a destination vector via Golden...summary on the Fragmid website.   Step 3 is simply a review of the assembled plasmid. It shows you...
  4. Viral Vectors 101: Calcium Sensors

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    Blog Post
    ...CEPIA (additional versions are available to simultaneously measure calcium changes in the ER, cytosol ...single fluorescent protein (FP), requiring only a simple measurement. However, single wavelength measurements...fluorescence to a red fluorescent state only by simultaneous (1) user-controlled illumination with violet...
  5. Corralling and Cornering the Goldilocks Antibody

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    Blog Post
    ...same target to see if the results are the same/similar, emphasizing the antibody’s specificity. Strength...from the background noise. If the sample images similarly to the negative control, that informs us that ...doi.org/10.1038/518027a Freedman LP, Cockburn IM, Simcoe TS (2015) The Economics of Reproducibility in Preclinical...
  6. Plasmids 101: The Wide World of Natural Plasmids

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    Blog Post
    ...by daughter cells during cellular reproduction, similar to the host’s chromosome, or they can spread between...plasmids likely have undiscovered functions, many are simply genetic parasites. Cryptic plasmids tend to be ... R., Sailland, A., Auriol, P., Vogel, T. M., & Simonet, P. (2001). Laboratory-scale evidence for lightning-mediated...
  7. Plasmids 101: Terminators and PolyA signals

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    Blog Post
    ...secondary structure to induce transcript cleavage, similar to the Rho-independent termination found in prokaryotes...terminate transcription. Polymerase I uses a process similar to the prokaryotic Rho-dependent mechanism, whereas...Definition of the upstream efficiency element of the simian virus 40 late polyadenylation signal by using in...
  8. Truncated gRNAs for Regulating Gene Expression

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    Blog Post
    ...bioengineering, as it has facilitated the development of a simple and scalable means of making targeted genetic edits...for instance, the challenge of attempting to simultaneously and selectively induce both cutting and regulation... targeting a different set of loci to induce simultaneous cutting and transcriptional activation or repression...
  9. Academic vs. Industry Postdocs

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    Blog Post
    ... Where publication is expected, the process is similar to academia. The only difference is that your paper...postdocs and do not support or manage them well are simply no longer allowed to have postdocs. In academia...could be laid off while you’re there. Of course, similarly, an academic advisor could also lose their funding...
  10. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

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    Blog Post
    ...biotinylated by BirA. From here, the CAPTURE method is similar to other 3C methods: crosslinking captures chromatin...well-characterized beta globin locus. The two methods had similar rates of specificity for the DNA sequence targeted...multiplexing and high resolution allowed the Xu Lab to simultaneously characterize the ‘social networks’ of 157 super...
  11. Degrading DNA with Cascade-Cas3

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    Blog Post
    ...perform its helicase and nuclease functions simultaneously, by continuously unraveling dsDNA, pulling ...reeling of ssDNA and unwinding of dsDNA occur simultaneously, which creates a loop in the target strand ...is not as well understood but likely occurs in similar regions. This combination of chewing up one stand...
  12. Addgene's Cancer Collection Pages Connect the Research Community

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    Blog Post
    ...the accelerator of a vehicle being stuck down. Similarly, a mutation that turns a tumor suppressor “off...cell to undergo programmed cell death while simultaneously stimulating growth and division. The combined...pathway. Its major appeal is that it enables the simple construction of vectors to study the function of...
  13. MXS Chaining

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    Blog Post
    ...constructs robustly, quickly, and simply. We hope that you can similarly apply MXS chaining to your experimental...Well suited for the modular assembly of highly similar or repetitive sequences. Intermediate chaining ...
  14. Plasmids 101: Secondary Nanobody Toolbox

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    Blog Post
    ...Nanobodies can be labeled with up to 3 dyes, yielding similar signal strength to a standard IgG secondary antibody...two-step staining with antibodies. Nanobodies also simplify multicolor staining experiments because they don...with this multi-color staining workflow yields similar localization patterns as cells stained for one ...
  15. Transferable Skills Guide: Identifying Your Transferable Skills

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    Blog Post
    ...and I guarantee you have developed them through similar experiences in graduate school. These skills will...they use, you’ve gained experience working with similar protocols, and look - they’re seeking someone who...strength until it is identified by another person. Similar reasoning may also apply to other acquired skills...
  16. Viral Vectors 101: The Retroviral Lifecycle

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    Blog Post
    ... we will mostly use the gamma-retroviruses as a simple spokesmodel for the whole family; however, you ...polymerase gene may seem like a mystery. But it’s simple. The RT protein is made by a separate set of instructions... a tool is done, and the transgenic genome will simply express from whatever promoters were included in...
  17. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

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    Blog Post
    ... filters you do not need to start a new search. Simply click the ‘X’ next to any filter to clear it. ... Tags: Identify a specific marker protein Use: Similar to “Vector Type” you can filter by experimental... plasmids that may be difficult to define for a simple search. These pages provide helpful information...
  18. AAVs CREATed for Gene Delivery to the CNS and PNS

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    Blog Post
    ...increase transduction efficiency, none allow for simple and efficient transduction of both the CNS and ... not lost in a jumble of fluorescent neurons. A simple but flawed approach to multi-color labeling is ...PubMed PMID: 28671695. 2. Deverman BE, Pravdo PL, Simpson BP, et al. Cre-dependent selection yields AAV variants...
  19. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

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    Blog Post
    ...minigenomes are incapable of producing infectious virus (similar to lentiviral vectors) because they do not encode...components of the Zaire ebola virus minigenome system Similar to other RNA viruses, EBOV brings its own RNA-dependent...studied independently of other viral proteins simplifying the interpretation of results. This system can...
  20. The Importance of a Fun Workplace: Company Culture at Addgene

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    Blog Post
    ...done annually - they are fan favorites! 6. Keep it simple This may be an obvious one, but don’t make things...really! We've had plenty of events that included a simple corn hole set and a cooler. Chances are one of ...company’s intranet. Don’t forget about the power of simple word of mouth - be sure to bring up your event ...
  21. Deciphering the Mysteries of Behavior with Viral Vectors

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    Blog Post
    ...less risk averse. Similar to the elevated plus maze, the light-dark box is a simple device comprised of...downregulating expression of miR124a achieved a similar improvement in the forced swim test.  Another brain...
  22. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

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    Blog Post
    ...transgenic zebrafish are well established (Felker and Mosimann, 2016). One common and robust strategy uses a ...tail. Compared to the opaque mouse embryo, where similar processes occur over days inside the mother, the... Central PMCID: PMC6247916. Felker, A., and C. Mosimann. "Contemporary zebrafish transgenesis with Tol2...
  23. Save Time with Transient Plant Leaf Transformations

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    Blog Post
    ...tobacco, Agrobacterium infiltration into leaves is a simple and efficient way to overexpress a gene and evaluate...abaxial site (lower surface) of the leaf. This is a simple and easy technique to implement. Success might ... Li, Jian-Feng, et al. "The FAST technique: a simplified Agrobacterium-based transformation method for...
  24. New Tools Enable CRISPRa for Neuroscience Applications

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    Blog Post
    ...sgRNAs. Its RNA-guided nature makes CRISPRa much simpler to implement in genome-wide screens over cDNA libraries... must be screened to identify effective ones. Simplified in vivo delivery using SPH mice CRISPRa has largely...PMCID: PMC6412672. Zhou, Haibo, et al. "In vivo simultaneous transcriptional activation of multiple genes...
  25. Hot Plasmids - August 2020

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    Blog Post
    ...//doi.org/10.1101/160374 Nanobodies- there's a simple method for that! By Michelle Cronin The benefits...Kimiko Hara, and Kazuo Umemura have described a simple method which alleviates the inclusion body problem...problem and makes nanobody production in E.coli as simple as following a DNA miniprep protocol!  First, the...
  26. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

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    Blog Post
    ...Just like analysis pipelines for DNA that group similar sequences together in bins, we thought that binning...mentor, as well as having a cohort experiencing similar situations to themselves, allowed the participants...greater effort in the future to match people within similar fields (ASM Microbe has eight tracks), something...
  27. New CRISPR Tools: Cas7-11 and PASTE

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    Blog Post
    ...identify a large variety of related bacteria with similar Cas systems. Some of these Cas systems have enzymes...of Cas7-11 were reported as up to 85%, broadly similar to shRNA and Cas13 systems in a range of mammalian... matter – the mutated copy could be replaced by simply dragging and dropping in a functional copy of the...
  28. Transferable Skills Guide: Creativity

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    Blog Post
    ...implemented a policy of never rejecting an idea simply because it's not how things were done in the past...’t have to be perfect or even good. The goal is simply to throw out ideas.  When your five minutes are...If you’re in a time crunch, your break can be as simple as a few minutes of meditation or a walk to the...
  29. Antibodies 101: Immunoprecipitation

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    Blog Post
    ...Immobilization is the key to immunoprecipitation and simply refers to the process of anchoring an antibody,...applications Oftentimes, an IP-appropriate antibody simply isn’t available for a specific target. In cases...binding proteins involved in histone modification. Similarly, RNA IP (RIP) can be used to isolate proteins ...
  30. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

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    Blog Post
    ...pBAD-bARGSer-AxeTxe, gas vesicle expression in E. coli is easy: simply grow cells in Lennox LB (10 g/L tryptone, 5 g/L...improved purity and contrast. For in vivo experiments, simply induce injected engineered cells via regular intraperitoneal...a chemical transfection reagent (e.g. PEI-MAX). Simply cotransfect a mixture of 1248 ng of CMV-gvpA-IRES-BFP...
  31. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

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    Blog Post
    ...for these faults. However, it’s still unclear if similar pitfalls exist for CRISPR screens and how best ...off-target cutting patterns in genome-wide screens similar to patterns seen in single-guide studies? Many ... Short gRNAs have lower off-target cutting and similar on-targeting cutting as full-length gRNAs Lastly...
  32. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

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    Blog Post
    ... and Sadowski 1985). The FLP system works in a similar fashion to the Cre system. The FLP recombinase ...excision, inversion, and translocation of DNA in a similar fashion as between loxP sites (For details see ...codon optimization resulted in FLPO, which shows similar efficiency to Cre (Kranz et al. 2010). These differences...
  33. Plasmids 101: Control Plasmids

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    Blog Post
    ...transfected into human cells. From this result one could simply conclude that the shRNA didn’t work, as the expression...level of Gene X in cells treated with Plasmid A is similar to the expression level of Gene X in the untreated... controls are negative controls that produce a similar product, but do not target an endogenous gene in...
  34. Data Freedom: The Expansion of Data Sharing in Research Publications

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    Blog Post
    ...efficiently trawled their own data? A distinct (perhaps similarly impoverished lab) might apply different or novel...although it might be argued various authors have simply chosen to submit elsewhere). The majority of the...types? And how long should data be archived? Simplifying data access Some of the above mentioned issues...
  35. Plasmids 101: The Promoter Region – Let's Go!

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    Blog Post
    ...between cell types or organisms, promoters must be similarly variable. Bacterial promoters only work in prokaryotic...related species from which they were derived. Similarly, the various eukaryotic cell types (mammalian,...expression mRNA Mammalian expression promoter from the simian vacuolating virus 40 Constitutive  May include ...
  36. Adeno-associated Viruses (AAVs) for Genome Editing

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    Blog Post
    ...edit your gene of choice. These vectors are very similar, both with polylinkers for the addition of gene-specific...at once. Therefore, applications that require simultaneous modification of all alleles of a gene, or which...which require modifying multiple genes simultaneously in the same cell, are more appropriate for a CRISPR-based...
  37. The Fluorescent Vegetables in Aptamer Soup

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    Blog Post
    ...a miniaturized version of Spinach that retains similar levels of fluorescence when bound to DFHBI despite...of artificial guide RNAs that can be used to simultaneously target cellular RNA to specific intracellular... Broccoli are derived from Broccoli, and share similar properties to Broccoli despite binding to a different...
  38. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

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    Blog Post
    ...fastest test that has been developed so far.  A similar method has been optimized to detect the SARS-CoV...and will not give a positive result for highly similar viruses. Unlike Cas12a, Cas13a does not require...illuminator or UV light illuminator. Approach: Highly similar to the SHERLOCK diagnostic test, this test varies...
  39. Transferable Skills Guide: Conflict Resolution

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    Blog Post
    ...confrontational situations with others because you simply cannot see things from their point of view and ...herself to the nearby village (and to the reader) by simply listening. Chances are, you know someone like this...how previous iterations of those teams resolved similar issues in the past, or how completely different...
  40. Filming Science Videos in the Age of Social Distancing

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    Blog Post
    ...Magic Pocket Cinema Camera 4K. It’s wonderful and simple to use, but when you need to juggle filming and...perfect height (Fig. 2). We recorded video using a simple, external video recorder and a mic clipped to her...video conferencing software. Interviewees could simply boot up Zoom and record themselves talking to their...
  41. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

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    Blog Post
    ...the immune response. The second strategy is the simple removal of the HIV-1 genome from infected cells...efficiency differed from 48-100% between strains. In a similar experiment, CRISPR reduced the HIV-1 copy number...evidence that an HIV-1 cure may be within our reach. Similar studies have shown that CRISPR can be used to combat...
  42. Thoughts on the Future of Research 2015 Boston Symposium

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    Blog Post
    ...postdocs. Unfortunately, even when universities have simple postdoc numbers sorted out there can be gaps in...important take-away from this session was the idea that simply satisfying a percent requirement for diversity ...difficult question but not one that should be abandoned simply because it is difficult and is key to the continued...
  43. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

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    Blog Post
    ...complementary Flpbow construct that is functionally similar to Cre-based Brainbow, but is controlled by FLP...-Veenboer C, Barker N, Klein AM, van Rheenen J, Simons BD, Clevers H. Cell. 2010 Oct 1;143(1):134-44. ...Hampel S, Chung P, McKellar CE, Hall D, Looger LL, Simpson JH. Nature Methods. 2011 Mar;8(3):253-9. doi: 10.1038...
  44. Antibodies 101: The Basics of Western Blotting

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    Blog Post
    ...cells? Immunoblotting or simply the western blot, or western, is one of the simplest methods to detect the...could contain unintended cross-reactive proteins. Similarly, if you vary the experimental conditions from ...
  45. Generating Mouse Models Using CRISPR/Cas9

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    Blog Post
    ...the go-to choice for studying mammalian biology. Similar to any model, mice are not without their problems...manipulate the mouse genome using CRISPR/Cas9. Similar to the human genome, the mouse genome is made up... finally, we have a technology, CRISPR, that is simple in concept, straightforward to use, and robust ...
  46. Finding Your Perfect Job After University

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    Blog Post
    ...I changed jobs. I highly recommend taking on a similar position in your field of interest before you dive... solution attempted was more meetings, but this simply seemed to create more bureaucracy. As with any ...anticipated and there can be a glut of individuals with similar skill sets. This is not to say that all commercial...
Showing: 201 - 250 of 649 results