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We narrowed to 604 results for: abo.2

Showing: 481 - 520 of 604 results

  1. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...backbone, which expresses the gRNA from a Drosophila U6:2 promoter and Cas9 from the actin 5C promoter. Addgene...backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors then used GMAP assembly...engineering (MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate multiple single-strand...number of tandemly expressed gRNAs respectively) and 2) the size of the deletion. Smaller deletions generally...but be sure to let us know if you'd like to write about your plasmids in a future blog post. No time to ...pooled libraries, but thanks to a deposit from the laboratory of Dr. Ji-Long Liu at Oxford University, we are...you plan to use GMAP for your experiments, think about depositing your new plasmids with Addgene to help...

  2. Hot Plasmids - May 2022

    Type
    Blog Post
    ... those that also infect the liver.    Fig. 2: Summary of the generational evolution of the AAV ... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here. ...

  3. Hot Plasmids - February 2022

    Type
    Blog Post
    ...acceptable variations in DR sequences. Figure 2: comparison of Cas 9 vs. Cas 12a expression cassettes... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.  ...

  4. Antibodies 101: Buffers, Storage, and Conjugates

    Type
    Blog Post
    ...with abundance whenever possible.      Fig 2: Try to inversely pair fluorophore strength with protein...fluorophore for an experiment is easy, but what about those times you need more than one?   Multiplex ...

  6. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    ...and are considered “high copy”. A fragment from the 2 micron circle (a natural yeast plasmid) allows for...to high copy numbers. This creates a potential metabolic burden on the yeast cells. Some phenotypes may...

  7. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    ...this year. Designing the donor plasmid Figure 2: Donor plasmid design strategy. A donor plasmid contains...the Addgene Blog Learn More about Homology Directed Repair Learn More about Non-Homologous End Joining .... For a C-terminal tag, as shown in the example above, we inserted a linker and GFP tag at the end of ... and 1kb of homologous DNA flanking both sides (about 2.7kb total)—is inserted into a plasmid backbone... Plasmids, plasmids everywhere To find out more about gene edited hiPS cell lines generated using the ...line. The editing strategy and design described above can be used to create similar donor plasmids for...microhomology-mediated end-joining (MMEJ). To find out more about gene edited hiPS cell lines generated using these...

  8. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ...Antibody Fragments. Antibodies (Basel). 2019 Apr 9;8(2):28. doi: 10.3390/antib8020028. PMID: 31544834; PMCID... time, when someone speaks about an affinity reagent, they’re talking about a standard antibody: An ~150kD...mutation, allowing diabodies to be used for assembling protein nanostructures. Diabodies readily co-crystalize...for clinical applications, including imaging.  Diabodies are ~60 kDa proteins formed by two Fab fragments...2019). Structural diversity and flexibility of diabodies. Methods. 10.1016/j.ymeth.2018.09.005 Muyldermans...

  9. Addgene: The First Twenty Years

    Type
    Blog Post
    ...beginning Let’s take a quick flash back to 2004. “Shrek 2” was the most popular movie in theaters, the Razr ...20th anniversary was about all the amazing years behind us, as much as it as about the amazing years in...Lunenfeld-Tanenbaum Research Institute, posting about Addgene’s 20th anniversary on Twitter. In the beginning...Julien Béthune, Professor at HAW Hamburg, posting about his Blue Flame Award on Bluesky. In 2016, we also... As the pandemic slowed, Addgene began to think about how we could expand our support for the research...part of the Neuroscience AntiBody Open Resource (NABOR) platform, supported by an NIH BRAIN Initiative....

  10. Seven Tips for Using LinkedIn as a Scientist

    Type
    Blog Post
    ...the "Contact info" link right below their photo. 2) Personalize all connection requests LinkedIn is a...include a note about where you met IRL (in real life) and perhaps what you talked about (or how good the... this rule, but not that many). I’m asked a lot about requesting connections to complete strangers for...have been able to tell when some start-ups were about to be bought by seeing certain groups of people ...using social media as a scientists Read blog posts about networking ...

  12. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    ...main decisions to make: 1. Which modification? and 2. Which viral vector should be used? Modifying gene...general consideration for behavioral testing in a laboratory, a high level of experimental control is required... after stereotaxic injection. Find more details about choosing AAV or lentiviral vectors in our previous...it’s crucial to appropriately time the injection about two weeks before the behavior test. This time window... Viral Vectors Resources on Addgene.org Learn about Addgene's viral service Find viral protocols and...

  13. How to Prepare for an Industry Interview

    Type
    Blog Post
    ...company culture.  It’s also a good idea to prepare 1-2 unique questions for each interviewer by looking at...you do to get ready for this interview? Maybe the above scenario sounds familiar to you. It was my experience...positions often required a one-hour presentation about a prior research project. In total, the interviews... find lists that have lots of general questions about job responsibilities, performance evaluations, professional...require. Frequently they will ask you to present about your prior research. This is a chance to highlight...

  14. Antibodies 101: ELISA (Enzyme-linked Immunosorbent Assay)

    Type
    Blog Post
    ... adapted to a competitive format. Figure 2: Overview of using Competitive ELISA to quantitatively...blog Learn about the advantages and disadvantages of using monoclonal antibodies. Read about other techniques... require two different antibodies. As mentioned above, it can also be used to characterize antibodies ...concentration of proteins in an ELISA well As mentioned above, the measurement of the concentration of proteins...

  15. RNA Extraction Without A Kit

    Type
    Blog Post
    ...156–159. https://doi.org/10.1016/0003-2697(87)90021-2 Chomczynski P, Sacchi N (2006) The single-step method...more foolproof (still without a kit!) As mentioned above, working with RNA requires keeping your samples ...lysis. This can be challenging depending on your laboratory situation or tissue collection method, so biotechnology...supernatant is extracted with chloroform as in the above no-kit method.  Researchers have also developed ...

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

    Type
    Blog Post
    ...inside the cell,” says Pedre Pérez. Figure 2: Micrographs of the biosensor expressed in the human...students as we have done in the past. Below, read about the five winners and their research.   Abhi Aggarwal...work has only been made possible due to the collaborative and highly interactive environment that Janelia...biosensors or biosensors that detect new redox metabolites are vital for researchers to better understand...

  17. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    ...strain (which will be covered in more detail in part 2 of this blog series) controls the cell types in which...blog Find our CRISPR blog posts Read blog posts about using Cre-lox Read our genome engineering blog posts... posts Resources on Addgene.org Find more about Cre-lox Read our Genome Engineering Guide Learn more...

  18. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...neuron-astrocyte, and cancer-stromal cell pairs.   Figure 2: Schematic of the interaction between the sender cell...s chromatin status into account (Fig 1a). The laboratory of Karmella Haynes at Emory recently contributed...custom design their own engineering plasmid tools in about a week. Taking advantage of Golden Gate cloning,...

  19. To Each HIS Own

    Type
    Blog Post
    ...take care when using this method.      Fig 2:  A, The polyhistidine tag is a string of histidine...that this blog has answered all of your questions about polyhistidine tags. For more antibody-related information...

  20. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...protocols 4.4 (2009): 495-505. PubMed PMID: 19300443. 2. Geraerts, Martine, et al. "Comparison of lentiviral...drawback of this approach is that it can be quite labor intensive. Target cells are transduced with serial...

  21. 7 Simple Strategies to Resolve Conflicts with Difficult Supervisors

    Type
    Blog Post
    ...out for help so you can get the support you need. 2) Prepare an agenda for every meeting Supervisors are... I heard about supervisors in all of the above categories. In addition, I also learned about two other...Complainers, who spend most of their time moaning about their troubles, Silent or unresponsive clams, ... students and postdoctoral fellows are hesitant about expressing their ideas, especially if their supervisors...it is interfering with your work. Avoid talking about your emotions, such as frustration or anger, because... like the problem to be resolved You know more about your work than your supervisor does. You can save...Listen to your supervisor’s viewpoint and brainstorm about mutually beneficial solutions Your supervisor might...

  22. Lentivirus ddPCR Titration

    Type
    Protocol
    ...Activation 95 10 2 1 Denaturation 94 0.5 2 40 Annealing/Extension 60 1 2 40 Enzyme Deactivation 98 10 2 1 Hold ...diploid cells, thus the reason for multiplying by 2. $$V = 2*{copies\ RRE \over copies\ RPP30}$$ Use the viruses...diluted 2-fold serially, the concentration of RRE positive droplets should decrease by a factor of 2 across...Lentivirus is generally considered biosafety level 2+. Please ensure that you are in compliance with your...channel pipette 200–1000 µL single channel pipette 2–50 µL multichannel pipette 20–200 µL multichannel ...Detach cells by incubating with 200 µL TrypLE for 1–2 min. Resuspend cells in 500 µL DMEM complete and transfer... DG8 cartridge into the cartridge holder. Using a 2–50 µL multichannel pipet, load 20 µL of the reaction...

  23. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...PCR tube Ice Bucket 2 μL Template DNA (10 ng-500 ng) 5 μl 10X Taq buffer with MgCl 2 1 μl dNTP mix (10 ... reagents on ice): 2 μL Template DNA (10 ng-500 ng) 5 μl 10X Taq buffer with MgCl 2 1 μl dNTP mix (10 ...normally sterile dH 2 O. To make a 100uM stock of any primer, add a number of µl of dH 2 O equal to the number...annealing temperature step-wise by 1-2°C. The rate of DNA synthesis is ~1-2 kb/min. The extension time can ...work adequately. Divalent cations such as Mg 2+ and Mn 2+ stabilize the buffer solution. These cations...specific instructions about extension time and temperatures. Initial Denaturation for 2 minutes at 94°C. Denature...(PCR). Basic PCR Program Initial Denaturation for 2 minutes at 94°C: This initiation step heats the double...

  24. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust GAL4...resources on the Addgene blog Learn more about SapTrap Read about CRISPR/Cas9 in C. elegans Browse our Plasmids...always emphasized the need for open science and collaboration. The field already has comprehensive reference...planned for August 15th, 2019. For more information about the aims and approach of CeNGEN, check out their...elegans. This database provides detailed information about C. elegans 3’UTRs structures, alternative polyadenylation...addition to the wealth of open resources described above, there were many plasmid tools presented at the ...by the worm community. For additional resources about C. elegans and the plasmids we have at Addgene check...

  25. Science communication: 9 Strategies to Get Your Foot in the Door

    Type
    Blog Post
    ...more tips on using social media as a scientist here. 2) Public engagement - Participate in science festivals...Sainsbury Laboratory. She is passionate about science communications and loves talking about science, ...Schmidt, a Marie Curie Fellow at The Sainsbury Laboratory. Are you a science student or early career researcher...break into science communication? Everybody goes about this in their own way. The career paths into science...Society gives training in media skills, writing about your research and residential communication. If ...

  26. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    ...non-specific binding in our IHC condition.   Fig. 2: Schematic of the goldilocks antibody selection process...Have you ever been stuck googling about an antibody that will be essential to your new research project...high-quality antibodies for IHC. Working with collaborators and support from the Brain Research Through ...extensive library of monoclonal antibodies; Addgene collaborates with these companies to create and provide general...pattern should align with what the atlases (mentioned above) and literature suggest, providing an expectation...open-source database, the mAb3D Atlas will foster collaboration, progress, and innovation by allowing scientists...

  27. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...systems to defend against both DNA and RNA viruses - 2 cas systems regulate at the DNA level and 1 at the... 2015 conference invited me to attend and speak about Addgene’s experience helping scientists share plasmids... with the first keynote presentation. He talked about the impact of the Human Genome project on its 25th...of the meeting was how much we still don’t know about RNA. One talk that really brought this home was ...presented by Todd Lowe from UCSC. Dr. Lowe talked about his work on the 500+ human tRNA genes and, until... Udi Qimron from Tel Aviv University then spoke about using CRISPR/cas to address the growing clinical...

  28. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    ...communicate the nuances of protein structure in a flat 2-D images. In the early days of structural biology,...Until then, you can only wonder what I am talking about. How do researchers use physical models of protein...students work closely with local researchers to learn about their research while creating physical models of...model is an effective teaching tool. You can read about Jacob’s initial reaction to this model on his blog...will wake up in the middle of the night dreaming about the conformational gymnastics that this protein ...Additional Resources on the Addgene Blog Learn All about CRISPR Use CRISPR for Genome Visualization Tips ...

  29. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ... already have an end product in mind, steps 1 and 2 will generally be straightforward. Likewise for step...directly back to your specific goal. When thinking about step 3, however, you may be surprised at the number...Delivery methods for mammalian cell lines As mentioned above, the expression system you choose in many ways dictates.... The user should review the current literature about their preferred model.  If your expression system... Check out our CRISPR Featured Topic Page Learn about CRISPR Delivery Using Ribonucleoproteins (RNPs) ...

  30. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    .... Find Plasmids from the publication at Addgene 2. Green, Alexander A., et al. "Toehold switches: de-novo-designed...allowed gene circuits to be deployed out of the laboratory in a cell-free and biosafe format that was stable...circuit-based sensors to be used outside of the laboratory. The detection threshold for these RNA sequences...sequences (3 nM), however, was quite high and above what would be expected in patient samples (e.g.: blood...

  31. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    ... to remove this default setting. Timesaving Tip #2: blastx searches are inherently slower than blastn...such as full plasmid sequences provided by the laboratories that deposit their plasmids with us or other...mismatches and gaps in the alignment. If you are curious about the differences in the blastn programs, check out...Inside Look at NGS Plasmid Quality Control Learn about our Snapgene-powered plasmid maps. Resources on...

  34. Hot Plasmids - October 2020

    Type
    Blog Post
    ...products have distinct localizations. Figure 2: Split-wrmScarlet labeling proteins with different... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's...fluorescent proteins coupled with Fibronectin intrabodies generated with mRNA display (FingRs), Xue Han...

  35. CRISPR in the Clinic

    Type
    Blog Post
    ... Cure for HIV/AIDS. Int J Mol Sci. 2023 Jan 13;24(2):1563. doi: 10.3390/ijms24021563 Resources on Addgene.org...trials The CRISPR therapies don’t stop with the list above! There are many, many more in the earlier stages... and others we just don’t have the time to talk about. Other treatments currently in clinical trials include...

  36. Personal Protective Equipment (PPE) for BSL-1 and BSL-2 Labs

    Type
    Protocol
    ... biosafety level 1 (BSL-1) and biosafety level 2 (BSL-2). The BSL-1 classification is for labs working...BSL-1 and BSL-2 labs. Protocols... Protocols Personal Protective Equipment (PPE) for BSL-1 and BSL-2 Labs Personal ...Personal Protective Equipment (PPE) for BSL-1 and BSL-2 Labs Intro to the Lab Bench Check out more protocols ...individuals from potential accidents such as spills. BSL-2 is different because it includes labs that work with...agents associated with diseases in healthy humans. BSL-2 includes all of the precautions needed in BSL-1, however...and/or face shields can be used as needed. For BSL-2 work always wear glasses/goggles in addition to the...

  37. A Look at Addgene's QC Process

    Type
    Blog Post
    ...contain the expected plasmid features.  Figure 2. Addgene’s basic QC process. Created with BioRender.com...results, and subsequent conclusions. Being vigilant about these errors is an important part of maintaining...

  38. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    ... Werner S, Marillonnet S. PLoS One. 2011 Feb 18;6(2):e16765. doi: 10.1371/journal.pone.0016765. PubMed...think about how much that’s on it are plants, and the animals you eat ate plants too. Think about how many...is Head of Synthetic Biology at the Sainsbury Laboratory, where she often feels more like an engineer ..., or pathogens to their hosts. I spoke with her about what motivates her research, the MoClo Kit she and...parts for plant synthetic biology Addgene: Tell me about the Golden Gate Modular Cloning (MoClo) Kit. What...make a 10-gene construct, you don’t have to worry about repeating elements anymore. The kit contains sequences...easy to express genes in, which makes it a good laboratory workhorse. Tips & tricks Addgene: Do you have...

  39. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    ...PMID: 27151215. PubMed Central PMCID: PMC4938170. 2. Zuris, John A., et al. "Cationic lipid-mediated delivery...scientists to make genomic alterations, bringing about a revolution in genome engineering, with new techniques...available by both the Jacob Corn and Alex Schier laboratories. Delivering Cas9-gRNA ribonucleoproteins  An...delivery of RNPs is electroporation (A in the figure above), which generates pores in the cell membrane, allowing...as lipid-mediated transfection (B in the figure above), are in the early stages of development. David ...proteins harboring receptor ligands (C in the figure above), which result in the internalization of Cas9-gRNA...show promise for in vivo studies, little is known about the immunogenicity and stability of the Cas9 protein...

  40. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...) 12 (>100) xCas9(3.7)-ABE 69 (1.4) 43 (2) 21 (3) Not examined 16 (>100) Average percent...Blog Learn about Engineered Cas9 PAM Variants Read about CRISPR Base Editing Learn about Homology Directed...editing specificity. We’re excited to learn more about xCas9 - here’s what we know so far! Why do we ...point mutations, also has a strict editing window about 15 nt upstream of the PAM. Even if there’s a PAM...in mammalian cells, finding xCas9 3.7 displayed about the same editing rate as SpCas9 at an NGG PAM. When...are exciting, there’s a lot we still don’t know about xCas9. “I’m not 100% sure xCas9 is going to be flat...Find CRISPR Plasmids for Mammalian Systems Learn about CRISPR History ...
Showing: 481 - 520 of 604 results