Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 809 results for: nin

Showing: 251 - 300 of 809 results

  1. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    Published
    April 16, 2024, 1:15 p.m.
    ...you’ll probably find lots of different plasmids containing the same protein of interest tagged with different...provide many advantages over fluorescent proteins, opening up new applications.   Figure 1: Live-cell...covalently linked to derivatives of its substrate O6-benzylguanine (BG) (Keppler et al., 2003). This engineered...FlAsH, ReAsH) Few Easy SNAP-tag 182 Benzylguanine (BG) Many Easy eDHFR 159 Trimpethoprim...Bottanelli et al., 2016) under CC-BY license. Combining orthogonal self-labeling tags on different proteins...able to get a plasmid ready-to-use without any cloning at all! As mentioned above, several variants of...

  2. Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!

    Type
    Blog Post
    Published
    July 18, 2017, 12:41 p.m.
    ...grant this year. These scientists work on projects spanning from the identification of plant compounds and...activities (1). She has developed a fast endophyte screening system and plans to employ the light‐inducible...integration vectors (see for example this yeast cloning kit) she intends to make yeast capable of desaturating... to bacteria and plants. Her research involves cloning cry genes from the soil bacterium Bacillus thuringiensis...bacterial expression system such as the pET GFP LIC cloning vector and the pOKD plasmids. The recombinant Cry...

  3. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    Published
    June 4, 2020, 1:22 p.m.
    ...glycosidic bond leads to the release of a purine (adenine or guanine) from the DNA. These apurinic sites if in...used for plasmid amplification are endA negative, meaning the gene has been removed/mutated to neutralize...without a kit Read our blog posts about plasmid cloning Browse our Plasmids 101 blog posts Resources on...Molecular Biology Reference Find protocols on plasmid cloning Find plasmids for your research ...

  4. 10 Ways to Share Your Science!

    Type
    Blog Post
    Published
    May 13, 2014, 8:34 p.m.
    ...Morehead Planetarium. Similar things are likely happening in your community, and, if they aren’t, think ...motivated more by the desire to help close the widening gap between science and society by connecting ...out how they got where they are. 8)   Look into training from an organization such as COMPASS. Although...their science matter. If you aren’t ready for a training or one isn’t available yet, order a copy of Nancy...University of California, Santa Cruz, has been turning scientists into confident and connected science...

  5. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    Published
    Nov. 25, 2013, 10:12 p.m.
    ...Horvath P, Moineu S, Mojica FJM, Wolf YI, Yakunin JvdO, Koonin EV. Nature Reviews Microbiology. 2011 Jun...powerful and adaptive defense strategy against life-threatening viruses. Evolving to survive Although the CRISPR...CRISPR/Cas adaptive immune system seems like the winning ticket to ensure prokaryotic survival; however,...Cong et al. and Mali et al. in January 2013 (7, 8). Nine months later, more than 1500 publications describe...

  6. Natural or synthetic? How Addgene’s dataset reveals trends in biological innovation

    Type
    Blog Post
    Published
    Nov. 8, 2018, 3:47 p.m.
    ...website in search of plasmids. Now, scientists are beginning to see Addgene as a large data set. Addgene has...database. Through a series of simulations and machine learning, they created a classifier that predicts whether...of 173 known synthetic and natural genes.  In designing and optimizing their algorithm, they found that...the genes deposited at Addgene were synthetic. But nine years later, over 20% of the genes deposited that...Nielsen, Alec AK, and Christopher A. Voigt. "Deep learning to predict the lab-of-origin of engineered DNA...

  7. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    Published
    Sept. 18, 2018, 1:33 p.m.
    ...halves (Fig. 1): one containing the microtubule binding domain and the second containing the growth promoting...number of fluorophores. You can find plasmids containing the cobalamin binding riboswitches used in the...developing a new imaging-based bacterial colony screening platform, Oliver Griesbeck’s lab set out to engineer...complexes are more efficient than the BE3 plasmid containing rat APOBEC1. Finally, they show that the ACE ...

  8. Science Careers: Unruly Interests Feed Many Paths

    Type
    Blog Post
    Published
    Sept. 26, 2017, 12:12 p.m.
    .... My eye was caught by a couple of unusual job openings, although they seemed too far afield at first ...forget: what would it be like? Then I spotted an opening for being an editor at Science magazine. That one...referees. And I am constantly teaching – often explaining a technical piece of research to others outside...of the world’s scientific discoveries into an entertaining podcast? Look at our multimedia producer and...Blog Posts Resources at Addgene.org Find Job Openings at Addgene ...

  9. Antibodies 101: Chimeric Antibodies

    Type
    Blog Post
    Published
    April 25, 2023, 1:15 p.m.
    ...great experimental plan that requires indirect staining of both targets in your specimen. You scour the...Why does isotype matter? Isotype matters when constraining using the indirect method since a secondary ...alternative isotype (purple, mouse). Depending on the cloning process, all constant regions, just the heavy chain...isotype can not be used for multiplexed indirect staining methods since the secondary antibody used will...specific antibody are cloned into a plasmid backbone containing all constant regions, just the heavy chain constant...

  10. Addgene's Top 10 Blog Posts of 2015

    Type
    Blog Post
    Published
    Dec. 7, 2015, 6 p.m.
    ... 101: Blue-White Screening Getting a lot of empty vector contaminants in your cloning experiments and ...Addgenie Jessica Welch explains how Blue-White screening could be a potential solution. 5. Cpf1: A New ...genomic locations. 6. CRISPR 101: Non-homologous End Joining So you know that CRISPR can be used to generate...gymnastics behind DNA repair by non-homologous end joining (NHEJ). 7. Career Coaching for Scientists: Why ...

  11. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    Published
    April 25, 2017, 2:30 p.m.
    ...Jellyfish (Aequorea victoria) GFP (wtGFP) by mutating Threonine 203 to Histidine (T203H) (1). wtGFP normally contains...performing several rounds of random mutagenesis screening for enhanced RFP variants. PA-mCherry (E26V / ...naturally occurring Dronpa protein (named after dron, a ninja term for vanishing, and pa, for photoactivation)...PALM, also FPALM) (11). For PALM, a small area containing inactive PA-FPs is photoactivated, imaged, and...and new variants are still being engineered. By combining PA-FPS with superresolution microscopy, scientists... Michelle Cronin...

  12. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...for p-azido-l-phenylalanine E. coli pEVOL-pBpF synthetase for p-benzoyl-l-phenylalanine E. coli pAcBac1..., 1978), a measurement that is unaffected by intervening proteins, lipid bilayers, organelles or other...due to tens of thousands of biomolecules each containing multiple binding crevices that can nonspecifically...labeling contrast can be improved using disulfide-containing compounds such as British anti-Lewisite (which...PMCID: PMC2802762. 2. Dumas, Anaëlle, et al. "Designing logical codon reassignment–Expanding the chemistry...

  13. With Our New Viral Service, We're Taking CRISPR Further!

    Type
    Blog Post
    Published
    Dec. 6, 2016, 3:30 p.m.
    ...produce custom gRNA-containing lentivirus, we are providing lentiviral preps containing highly requested ... lentivirus is not always an easy process. The cloning capacity of lentiviral vectors (~8.5 kb of insert...optimization-scale infection and at least one screening-scale experiment achieving the gRNA representation...expertise as we’ve gotten these services up and running. Both Alex Chavez and John Doench are members of...

  14. Summer SciComm Series: A PhD in Science Communication

    Type
    Blog Post
    Published
    Aug. 23, 2022, 1:15 p.m.
    ...cells I was using in an experiment, then spend the evening listen to some bloke at the pub telling me that...to wonder how he has survived this long without downing any dihydrogen monoxide (H2O). Studying science...the same abstract but with the names reversed, meaning that the findings of the study were also reversed...were exhibiting a phenomenon called “motivated reasoning,” the catchall term for a range of neat little...nclimate1547 Kunda, Z. (1990). The case for motivated reasoning. Psychological bulletin, 108(3), 480. More resources...

  15. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    Published
    May 3, 2018, 1:48 p.m.
    ...diversity is impressive, and researchers are just beginning to tap the wealth of CRISPR possibilities. To ... are described below. Yan et al.: WYL-domain-containing accessory protein WYL1 increases RspCas13d and...EsCas13d activity, indicating that WYL domain-containing proteins may be broader regulators of Cas13d ...Cas13d enzymes is another win for bioinformatic data mining. As we continue to harness the natural diversity...Effector Positively Modulated by a WYL-Domain-Containing Accessory Protein.” Mol Cell. (2018) pii: S1097...

  16. A Quick Guide to a Career in Software Product Management

    Type
    Blog Post
    Published
    Nov. 24, 2020, 2:15 p.m.
    ...at my company was open to meeting with me and explaining more of her role, which eventually led me to ... problems that they are trying to solve. After learning more about the stakeholders, the product managers...next-gen sequencing (NGS), and high-throughput screening. While some developers had science backgrounds...audience. Additionally, in order to keep everything running smoothly, you’ll want to be really organized. Often... hours ahead, so I’m always catching up in the morning! 9:00 AM--Technical implementation review. The ...

  17. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    Published
    June 6, 2023, 1:15 p.m.
    ...generally have moderate to high immunogenicity, meaning they induce an immune response in the host. So...required components are very similar. While plasmids containing virus-specific elements such as the LTRs or structural...allows it to replicate episomally, essentially remaining an extrachromosomal part of the host genome that...endogenous DNA. AAV also has low immunogenicity, meaning it does not induce the same high immune response...which ones could be helpful to you. Interested in learning more about a specific virus? We have you covered...

  18. Viral Vectors 101: Gamma-Retroviral Packaging Systems

    Type
    Blog Post
    Published
    Aug. 6, 2024, 1:15 p.m.
    ...particle containing Matrix, Capsid, and Nucleocapsid components. Pol Precursor protein containing Reverse...example: pLXSN has the wild type LTR, an MCS for cloning X gene, an SV40 promoter, and Neomycin selection...either side of a retroviral provirus (see below). Cloning capacity between the LTRs is ∼8.5 kb, but inserts... The transcript begins, by definition, at the beginning of R, is capped, and proceeds through U5 and the...

  19. Clear and Accessible Writing

    Type
    Blog Post
    Published
    Feb. 15, 2024, 2:15 p.m.
    ...years. But learn we did! From Addgene's very beginning, we had to consider digital accessibility for ...student using viral vectors for the first time and learning how to purify AAV.  Second, you’ll identify your...captured at the 17%, 25%, and 60% layers, and genome-containing virions are captured at the 40% layer. To collect...insert a syringe at the interface of the 40% layer containing the virus and 60% iodixanol layer, with the bevel...alt text in the comments! We learned a lot from listening to others' alt text descriptions, and we’d love...

  20. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    Published
    Feb. 25, 2014, 7:11 p.m.
    ... can be easily maintained when grown on media containing the missing nutrient. Scientists can exploit ...the host’s auxotrophy. When grown on media NOT containing the nutrient, the host cells will die unless ...weak. yes   LYS2 L-lysine yes - Grow on plates containing alpha-aminoadipate in the absence of a nitrogen...used in gene disruption experiments. MET15 L-methionine and overproduces hydrosulfide ions yes - Grow... L-leucine   S. pombe  no no   ade6+ purine (adenine)   S. pombe  no no   Considerations when using...

  21. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    Published
    Jan. 16, 2018, 3:20 p.m.
    ... labeling using a noncanonical amino acid, azidohomoalanine (AHA). In the technique, AHA is incorporated...incorporated into newly synthesized proteins instead of methionine. The labeled proteins can be detected using click-chemistry...click-chemistry that covalently links the azido containing AHA to a fluorescent alkyne tag. The newly synthesized...determine the turnover of a specific protein by combining it with the proximity ligation assay (PLA). The...concerts and bake. you can contact her at rojas@ti2learning.org. References 1. Dieterich, Daniela C., et al...

  22. Summer SciComm Series: Cognitive Load

    Type
    Blog Post
    Published
    June 16, 2022, 1:15 p.m.
    ...audience’s mental bandwidth.  Jargon Learning, using, and retaining new vocabulary takes time and effort...others. But sometimes we’re unaware that we are leaning on an unspoken expectation of our audience knowing...concept to another which leads us to… Framework Learning to think like an immunologist, a geneticist, a...needed to remove a distraction, such as pausing or turning off the slide deck (or reminding them to put their...

  23. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    Published
    Aug. 5, 2021, 12:15 p.m.
    ...hydropersulfide that is then transferred to thiol-containing acceptors. These hydropersulfides modulate protein... biosensor can also be used along other thiol-containing acceptors in competition assays. “I believe that...photoconvertible fluorescent protein mEOS4a. Cells containing a sensor that shows a favorable outcome are then... to engineer many types of biosensors.   This screening method allowed Tian to optimize a red‑shifted ...neural microcircuits. “In the longer term, this screening platform may help us to create high-performance...

  24. Addgene: The First Twenty Years

    Type
    Blog Post
    Published
    Dec. 19, 2024, 2:15 p.m.
    ...Addgene’s 20th anniversary on Twitter. In the beginning Let’s take a quick flash back to 2004. “Shrek ...business practices, like accepting e-signatures and designing a website users could easily navigate. This meant... make sharing reagents and protocols so easy!” — Nina Jain, Harvard Medical School. We want to give a ...requestors alike. We transformed this process by aligning institutions with standardized agreements and ...Addgene for the basics of molecular biology and cloning, since they have reasonable introductions to lots...

  25. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    Published
    Dec. 16, 2015, 3:30 p.m.
    ...How Non-Homologous End Joining Repairs Double Strand Breaks Get Tips on Designing Your gRNA Browse Our ...RuvC groove, Slaymaker et al. made a variety of alanine substitutions throughout the groove in 32 separate...diminish off-target cutting (4). However, instead of weakening interaction sbetween the non-target strand and... in applied and/or therapeutic settings. Will combining the different mutations found in each of these...

  26. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...produced in E1-containing mammalian cell lines such as HEK293 cells. 1st generation AdV cloning capacity is.... 2nd Gen AdVs were designed to have increased cloning capacity, decreased  potential for the generation...deleted in 2nd generation AdVs, thereby increasing cloning capacity to ~12 Kb. The ever popular pAdEasy system... interest for gene therapy due their increased cloning capacity (up to 36 kb!), long-term transgene expression...

  27. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...sequence of the plasmid, we examine three things: Aligning the NGS result to a reference sequence to confirm...backbone elements.   Confirming the gene/insert by aligning to NCBI entry or using BLAST.   Confirming tags.... We’ll break down each of these steps below. Aligning the sequence The first step we take when performing...publicly available reference sequence or plasmids containing genes with many modifications. We look for point...-Cas9 knockout and transcriptional activation screening. Nat Protoc 12:828–863 . https://doi.org/10.1038...

  28. CRISPR in the Clinic

    Type
    Blog Post
    Published
    Nov. 2, 2023, 1:15 p.m.
    ...treatment strategies for these disorders have involved turning on fetal hemoglobin to compensate for defects in...approach, editing patient cells ex vivo and then returning them to the patient. The first human trial involving...many, many pieces. Locus Biosciences is currently running this trial, with more updates expected in the coming...considerations for all CRISPR therapies, but especially concerning when using a systemic approach. Regarding effectiveness...difference, then the treatment will not work in a meaningful way. Delivery, especially for systemic approaches...

  29. A Look at Addgene's QC Process

    Type
    Blog Post
    Published
    May 6, 2025, 1:15 p.m.
    ...vigilant about these errors is an important part of maintaining the quality of reagents, especially shared ones... easy to edit, and easy to share. However, the cloning process — including the propagation of plasmids...these errors could cause the plasmid to stop functioning properly or be something completely different...process, we believe this to be non-negotiable. By maintaining a high standard of quality control, we have built... built a trusted plasmid catalog containing over 150,000 sequence-verified constructs to share with scientists...

  30. Engaging with science and society at pgEd

    Type
    Blog Post
    Published
    Sept. 20, 2018, 12:17 p.m.
    ...lesson plans that are made freely available online; running professional development (PD) workshops for teachers...organizing Congressional briefings in Washington DC; convening meetings of experts and leaders from academia,...number of different opportunities to broaden my training beyond the biological sciences, as well as to ...students. One of these organizations was pgEd. After obtaining my PhD and working in digital academic publishing...festivals, and getting experience in either giving or running these events will be important both in preparing...

  31. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    Published
    July 14, 2016, 2:30 p.m.
    ..., Terns MP, White MF, Yakunin AF, Garrett RA, van der Oost J, Backofen R, Koonin EV. An updated evolutionary...transfer, analogous to traditional restriction enzyme cloning. Sticky-end mediated gene transfer would be particularly...homology-directed repair (HDR) over non-homologous endjoining (NHEJ). Cas9-mediated NHEJ usually destroys the...Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin EV, Zhang F. Cpf1 Is a Single RNA-Guided Endonuclease...

  32. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    Published
    Feb. 4, 2020, 2:15 p.m.
    ...naked eye. This makes cloning quite a bit easier since you can see if your cloning experiment worked just...methods. While techniques such as  blue-white screening require X-gal, an expensive exogenously-added ...chromoproteins don’t require other substrates for screening. Thus chromoproteins can be used as markers in... between chromoproteins. In some cases, cells containing high-copy plasmid-borne chromoproteins lost color...

  33. Antibodies 101: Flow Compensation

    Type
    Blog Post
    Published
    May 16, 2024, 1:15 p.m.
    ... antibodies may have similar emission spectra, meaning they will emit fluorescent light at similar wavelengths...record the fluorescence for both at similar levels, meaning that your populations will ‘look’ similar on your...compensate in the top right corner of a flow plot, meaning that it should be double-positive for the two markers... what two markers your fluorescent markers are staining for. If a cell is positive for both markers, it...compensation values later. This can be helpful as you're learning! Ashlyn has a research background in CD8+ T cells...

  34. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    Published
    April 18, 2024, 1:15 p.m.
    ...collection of CRISPR tools has grown steadily, gaining approximately 2,000 plasmids/year from 2017 to ...to 2023. This growth is supported by new labs joining the Addgene CRISPR community, with approximately ...wanted to know, so we classified CRISPR plasmids containing a nuclease into three functional groups: nucleases...grown to 17,560 CRISPR plasmids. That’s almost a ninety-fold increase in size! From this collection, we...educational resources help scientists navigate that learning curve. As of 2023, our CRISPR guide has been viewed...

  35. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    Published
    Feb. 25, 2016, 3:30 p.m.
    ... of these resources as I could. Networking and learning from the careers of others were key to creating...productive partnership CINSO has had. Volunteer! The turning point in my graduate experience, the point at which...Dinosphere” almost every week. I gained experience explaining science to visitors of all ages, learned a lot...lessons they have learned during their scientific training and wrote about them in a blog post on Born to...to Science: Life Lessons from Scientific Training More and more voices of encouragement emphasize the...

  36. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    Published
    Sept. 24, 2015, 2:30 p.m.
    ...delivery methods.   General considerations for planning your CRISPR experiment As with any experiment,... factors that need to be considered during the planning process. For CRISPR experiments, the following...viral delivery. Cas9 mRNA and gRNA Plasmids containing gRNA and Cas9 are used in in vitro transcription...as well as genome repair by non-homologous end joining (NHEJ) or HDR. EGFP validation of sgRNAs: Can evaluate...evaluate component delivery and sgRNA efficacy by cloning in genome target sequence into EGFP reporter. Target...

  37. SciComm: Taking Science to Elementary and Middle Schools

    Type
    Blog Post
    Published
    July 9, 2019, 12:52 p.m.
    ...discretionary money. They also usually have a good event-planning base and will know how to reserve space, time ...themselves as “scientists.” We had some accountants running our math tables!  Be realistic about what you...you can teach  5-10 year olds in a single evening. Grades K-5 cover a broad range of attention spans, skill... simple. I honed the complexity of the talk by running it by the teachers and sharing some slides on Twitter...Even the most ambitious teachers, who work hard maintaining a current, dynamic curriculum, would be hard-...

  38. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    Published
    July 30, 2020, 1:15 p.m.
    ...detection had over 1,000 authors. The chapter on deep learning addresses the dilemma of creating a technology... “The most obvious future consequence of the undermining of the scientific process and the marginalization... the summer of 2020 that the consequences of undermining science are even more immediate and dire than...himself has written entire books about optogenetics. Defining science and summarizing its past, present, and...task to set oneself. As a scientist who enjoys learning a little bit about many different things (part...

  39. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ...plasmid cloning blog posts Resources on Addgene.org Find protocols for molecular biology and cloning Read... use a phenol chloroform extraction to remove remaining proteins and RNase. Finally, DNA is precipitated...into this mixture to absorb the nucleic acids. Containinants bind cellulose less tightly than nucleic acids..., as detected by qPCR, but this did not reduce cloning efficiencies.  When compared to fresh columns, ...

  40. Great Results Start with Great Standard Curves

    Type
    Blog Post
    Published
    Aug. 15, 2024, 1:15 p.m.
    ...standard in their bicinchoninic acid assay (BCA) to normalize samples before running a western blot. Standard...provide some general considerations for scientists planning to use them in their assays.  What is a standard... above 0.95.    Additional considerations When running assays that rely on a standard curve, it is critical...program. Summary Few things in the lab are more disheartening than seeing a potentially exciting result quashed...

  41. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    Published
    Dec. 7, 2020, 2:53 p.m.
    ... direct RNA-guided transposition. By natively combining the DNA integration capabilities of transposases...with Type I-F INTEGRATE using a single plasmid containing a multi-spacer CRISPR array. Workflow and...s41586-019-1323-z Peters JE, Makarova KS, Shmakov S, Koonin EV (2017) Recruitment of CRISPR-Cas systems by ...Ladha A, Gardner Z, Schmid-Burgk JL, Makarova KS, Koonin EV, Zhang F (2019) RNA-guided DNA insertion with...science.aax9181 Strecker J, Ladha A, Makarova KS, Koonin EV, Zhang F (2020) Response to Comment on “RNA-...

  42. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    Published
    Aug. 20, 2020, 2:30 p.m.
    ... genome. These off-target sequences may be unintentionally mutated along with the desired target locus...sequences when used in place of wild-type SpCas9. The remaining variants (VQR, EQR, and VRER) recognize novel ...Staphylococcus aureus (Sa) NGRRT or NGRRN Neisseria meningitidis (Nm or Nme) NNNNGATT Campylobacter jejuni...Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F (2015) In vivo genome editing...Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin EV, Zhang F (2015) Cpf1 Is a Single RNA-Guided ...

  43. Transferable Skills Guide: Creativity

    Type
    Blog Post
    Published
    Jan. 18, 2022, 2:15 p.m.
    ...identifies nine enemies that stand in the way of creative thinking. I won’t go through all nine here, but...found that many researchers feel they lack formal training in a variety of transferable skills. At Addgene...absolutely a transferable skill developed during PhD training. Creativity is required to develop new projects...’re stuck drawing stick figures, remember your training as a scientist has been building your creativity...

  44. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...checkpoint.  The plasmid is self-transmissible, meaning that the plasmid is conjugative and carries the...inducibility (Toh et al. 2023), and additional cloning sites (Keen 1988).  Find broad host range recombinant... Origin of the Broad Host Range Plasmid RK2: Positioning of Various Motifs is Critical for Initiation ...1988). Improved broad-host-range plasmids for DNA cloning in Gram-negative bacteria. Gene, 70(1), 191–197...R. (1988). Chapter 15—Broad-Host-Range Plasmid Cloning Vectors for Gram-Negative Bacteria. In R. L. Rodriguez...

  45. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    Published
    Oct. 13, 2022, 1:45 p.m.
    ...HypaCas9, are great choices. Check out our SpCas9 containing plasmids, optimized for a variety of species....critical enzymatic residues has been converted to an alanine (D10A or H840A), changing its double-strand cut...available. This system utilizes an RNA template containing the desired edit adjoined to the sgRNA, a reverse....   Ok, I have a Cas9! What do i do next? Designing or selecting a sgRNA Addgene may carry a validated...expression systems Addgene carries a variety of Cas9 containing plasmids that have been optimized for expression...

  46. Welcome to the Lab Bench! New Protocol Series Helps Trainees Get Started in the Lab

    Type
    Blog Post
    Published
    Sept. 7, 2021, 1:15 p.m.
    ... shift to remote learning due to COVID-19. We had many protocols for plasmid cloning and working with ...details about personal protective equipment to learning about how to use a tissue culture hood and other...on our Protocols page and YouTube page.  Happy learning, and welcome to the lab bench, new scientists!...

  47. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    Published
    March 5, 2019, 1:08 p.m.
    ...involves the introduction of a transgene construct containing fluorescent proteins (XFP) of different colors...not undergo recombination can be detected by immunostaining of the truncated YFP. In Thy1-Brainbow mice...are that it does not use ubiquitous promoters, meaning that not all cell types can be labeled, and that...labels Drosophila neurons at the neuroblast stage, meaning that neurons of the same lineage will express the... a plasmid construct called Zebrabow-GateDest containing an RFP, CFP, and YFP downstream of a ubiquitous...

  48. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    Published
    Sept. 29, 2020, 1:15 p.m.
    ...around 20 minutes, they are perfect for molecular cloning and metabolic engineering.  The E. coli genome ....  Check out Addgene’s collection of plasmids containing Rattus norvegicus genes! Mexican axolotl (Ambystoma...community.  The Arabidopsis genome project marked the beginning of widespread Arabidopsis genetics research development...out the GreenGate Toolkit which allows for easy cloning of constructs specifically designed for use in ...Xing, X.-H. (2018). Pooled CRISPR interference screening enables genome-scale functional genomics study...

  49. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ...been around since 1975 thanks to the Nobel prize winning work of Milstein and Köhler who developed hybridoma...by infecting bacteria for additional rounds of panning and scFv characterization.    Figure 2: ...by infecting bacteria for additional rounds of screening. Created with Biorender.com.   Some phage ...and VH sequences. Phage display also allows for screening scFv generated from the B cell antibody genes ...Gąciarz A, Ruddock LW (2017) Complementarity determining regions and frameworks contribute to the disulfide...

  50. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    Published
    May 20, 2021, 1:15 p.m.
    ...months. Jennifer and I spent about an hour that morning scrambling. Everything we had set up involved multiple...adapted for video making.   Cycle One: The Ironing Board Control Center With the pandemic in full ...that included iPad and her laptop raised on an ironing board, giving it the perfect height (Fig. 2). We...to think of something else.   Figure 2: Ironing boards. Good as a WFH desk, and as a video control...smooth filming in the future, but we were finally refining the process down to a place I could be happy with...
Showing: 251 - 300 of 809 results