We narrowed to 618 results for: abo.2

...perceived climate change risks. Nature Climate Change, 2(10), 732-735. doi:10.1038/nclimate1547 Kunda, Z. (...discussions about the pros and cons of different fabrics while we worked. We also had discussions about the difference...grad student. I chose to focus on misinformation about sexual and reproductive health, as the stigma around... misinformation can flourish. I’ve never forgot about all the anxious, whispered questions from friends...the study design or by their preconceived ideas about the controversial medical issues. Participants ...none of the participants had preconceived opinions about “adeolitis.” The participants in my study were ...make reusable cloth menstrual pads while we talked about the environmental impact of disposable products....

...and in line with Addgene style.        Fig. 2: Reference image (left) provided to our graphic designer...We also want to tell you about things you didn’t know you wanted to know about, like research breakthroughs... want to know more about! You can also suggest blog topics or even reach out about writing a guest post...technical skills, and experiences from thousands of laboratories that makes up our Help Center, blog, website...

...create a recombinant antibody.     Figure 2: Changes to the heavy chain or light chain genes within... Learn more about research applications for antibodies in Intro to Antibodies Read about the Developmental...antibodies in your experiments, you may be curious about all of the different kinds of antibodies that are...Disadvantages of monoclonal antibodies As mentioned above, genetic drift is one concern when using hybridomas...Hybridoma Bank, a repository for hybridomas  Learn about monoclonal antibodies for immunotherapy Resource...

...axon terminals to the soma (retrograde tracer) (Fig. 2). Viruses as neuronal scalpels Viruses can be injected... Get Started with AAV Learn more about Lentivirus Find out More about Addgene's Viral Service Resources...We often think about the brain’s function in terms of its regions. But equally important is the way the...head bone’s connected to the neck bone,” but with about a hundred billion neurons and six hundred trillion...

...to visualize multiprotein complexes.  Fig. 2. Multiplex immunolabeling with subclass-switched recombinant... called NABOR (Neuroscience AntiBody Open Resource). The antibodies distributed through NABOR are recombinant...troubleshooting methodologies drew me to learn more about the process that was used to create this important...secondary antibodies are commonly used in research laboratories to detect the different subclasses, but there...

...improve the brightness of previous Nano-lanterns by 2-4 fold and permit five-color fluorescent imaging! ...Since their first publication in 2012, the Nagai laboratory has assembled a collection of multicolored nano-lanterns...Nano-lanterns, Takai et al. continuously imaged cells for about 4 hours! The latest enhanced Nano-lanterns (eNL)... Nano-lanterns and your experiments The Nagai laboratory has a done a wonderful job characterizing Nano-lanterns...Additional Resources on the Addgene Blog Learn about Photoactivatable Fluorescent Proteins Check Out ...

...work with Grows extremely fast (doubling time of ~ 2 hours) Amenable to genetic manipulation The first...pombe have dominated the research scene. But what about the other yeasts – in the biomedical world, are ...pharmaceutical companies (along with some research laboratories) have picked up the system for some of the uses...eukaryotic cells and peroxisomes are where methanol metabolism occurs in methylotrophic organisms. K. phaffii...

...Without binding to the guide RNA, Cas9 cannot cut. 2. Bind to target DNA in the presence of a guide RNA...that you know about all the different components of CRISPR, It's time to start thinking about how you can...Use the following flow chart to get you thinking about the types of experiments you're planning on doing...

...two options to begin your plasmid deposit. Figure 2: Choose to submit plasmid(s) online or by using a ...format. Figure 6: Add sequence, maps, and files. Step 2: Gene and Insert *Note: This page will not appear ...annotate on this page. Figure 7: Enter information about the gene/insert(s). Step 3: Cloning Information ...

...responsible for that differentiation.   Fig. 2: Single cell RNA-seq clustering of cells transduced...changes. Using this road map, and what was known about previously characterized TFs, the Zhang group were...Additional Resources on the Addgene Blog Learn More about Lentivirus Figure out How to Get the Most from Your...

...phosphate monobasic monohydrate (NaH 2 PO 4 ∙H 2 O), pH 7.0 138 g NaH 2 PO 4 ∙H 2 O 1 L deionized water Adjust...sodium phosphate dibasic (NaH 2 PO 4 ), pH 7.0 142 g of NaH 2 PO 4 ∙H 2 O 1 L deionized water Adjust pH...monobasic monohydrate (NaH 2 PO 4 ∙H 2 O) 610 mL of sterile sodium phosphate dibasic (NaH 2 PO 4 ) Store up to...recombinant antibody. Section 2: Buffer exchange Choose Option 1 or Option 2 based on the concentration ...antibodies. Workflow Timeline Day 1: Purify antibody Day 2 or later: Buffer exchange Equipment Class II, Type...conical tubes NanoDrop spectrophotometer 37 °C, 5% CO 2 incubator with shaking platform set to 120 rpm 37 ... channel pipette 20–200 µL single channel pipette 2–20 µL single channel pipette Reagents Aspirating pipette...

...retransduce a model organism such as mice, e.g. first at 2 months, then at 4 months? Or do antibodies become ...labeling in the brain with PHP.eB: A recent CLOVER collaboration injected 1e+13 vg per mouse to get phenotypes...coverage. Please see this paper for more information about PHP.eB brain labeling at various doses.  As always...

...account when simulating a digest as depicted in Lane 2. Controlling methylation Finally, you can control...questions is the same--methylation! Read on to learn about how DNA methylation may affect your restriction ...be methylated. Using the same example plasmid as above (which has the XbaI site blocked by methylation)...

... tabular HTML formatted output. Surprisingly only 2 tools provide FASTA output. A few notable tools allow...Nature biotechnology(2014). PubMed PMID: 24880488. 2. Hsu, Patrick D., et al. "DNA targeting specificity...databases mentioned above or, even better, first-hand knowledge from laboratories employing CRISPR on ... CRISPR system was involved. Today, that same laboratory enjoys a successful edit rate of over 90% in ...targetable NGG PAM sites whereas the human genome has about 300 million. With such a sparsely targetable genome...worse by an ever increasing marketplace. Since 2013 about 11 tools have been added every year.   Breaking ...databases are still young and will require a collaborative effort from the community in order to succeed...

...cellular physiology.  https://doi.org/10.1002/jcp.30064 2. Barman et al. 2020. “A glance at genome editing with...therapeutic.” There are many fundamental questions about CRISPR/Cas biology still left to answer as well,...

...retroviral library was used to transduce T helper type 2 (Th2) cells from mice that constitutively express ... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.    ...

... enrich the microvesicles containing AAVs (Figure 2). Since the development of the exo-AAV isolation protocol...vectors 101 eBook Resources on Addgene.org Learn about Addgene's viral service Find viral protocols and...

... 293HEK Inhibits termination step of translation 2-10 ug/mL G418/Geneticin neo HeLa, NIH3T3, CHO, 293HEK...within 3-5 days and resistant colonies appear in about 10-14 days depending on how quickly your cells divide...

...430825, 175 cm 2 Cellstack 5, Corning 3319, 3180 cm 2 Cellstack 2, Corning 3269, 1272 cm 2 Heat-inactivated...: 50 mM Tris HCl, 150 mM NaCl, 2 mM MgCl 2 Add the following to the 2 L sterile bottle: 1836 mL deionized... solution can be stored at 4 °C for up to 2 months. After 2 months, discard the tube and thaw a new working... mL of PBS. Aspirate PBS and add 2 mL of 0.05% Trypsin/EDTA. Wait ~2 min. Neutralize trypsin by adding...Cell-Stack (CS5) (Link opens in a new window) (3,180 cm 2 - the same surface area as 21 x T-175 flasks). Cell...flasks. Workflow Timeline Day 0: Seed cells in CS2 Day 2: Seed cells in CS5 Day 3 (am): Transfect cells Day...Biological Safety Cabinet 0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel...

...materials D.2 Screening for inserts E. Producing Lentiviral Particles E.1 Recommended materials E.2 Protocol...Published articles H.2 Web resources I. Appendix I.1 Sequence of pLKO.1 TRC-Cloning Vector I.2 Recipes I.3 Warranty...VWR: #7177-48-2. Use at 100 μg/mL. Carbenicillin VWR: #80030-956. Use at 100 μg/mL. C.2 Annealing Oligos...oligo 5 μL Reverse oligo 5 μL 10x NEB buffer 2 35 μL ddH 2 O Incubate for 4 minutes at 95°C in a PCR machine...buffer 1 1 μL AgeI add ddH 2 O to bring to 50 μL final volume Incubate at 37°C for 2 hours. Purify with Qiaquick...buffer for EcoRI 1 μL EcoRI 14 μL ddH 2 O Incubate at 37°C for 2 hours. Run digested DNA on 0.8% low melting... For a standard T4 ligation, mix: 2 μL annealed oligo from step C.2 20 ng digested pLKO.1 TRC-cloning ...

...types within a sample to expected labeling (Figure 2). The panel of antibodies used in this approach should...the antibody in the same tube.     Figure 2: Schematic showing the use of an orthogonal approach...leukocytes. The file shared also contains information about the labeling pattern of several cell lines using...Laflamme, C., Virk, H., NeuroSGC/YCharOS/EDDU Collaborative Group, & ABIF Consortium. (2024). A guide to...

...the details and learn more about colony PCR in this blog post.   Figure 2: The steps of colony PCR...often relies on antibiotic resistance. But what about identifying clones that contain your desired insert... restriction digest. The key is to be strategic about your enzyme choices. You’ll want to pick enzymes...Sanger sequencing or the other techniques mentioned above. In molecular biology, cloning a gene into your ... and tips Resources on Addgene.org Learn more about Sanger sequencing Read the Molecular Biology Reference...

...PubMed PMID: 8491198. PubMed Central PMCID: PMC413431. 2. Bukrinsky MI, et al. “Active nuclear import of human...applications to help you start (safely!) thinking about the possibilities. Ready to plan your next (or maybe...S, Varmus H. Retroviruses. Cold Spring Harbor Laboratory Press; Cold Spring Harbor, NY, USA: 1997. PubMed...Additional Resources on the Addgene Blog Learn More about Lentivirus Listen to Connie Cepko Discuss Gene Therapy...Addgene.org Browse Our Viral Vector Pages Learn About Viral Vector Biosafety Deposit Your Viral Vectors...

...JS, Legube G, Miller KM Genes Dev. 2015 Jan 15;29(2):197-211. DNA Cross-Bridging Shapes a Single Nucleus...Leah Bury of Microscopic Art. I started thinking about this problem when I recently attended the 2018 Chromosome...lot of scientists who not only remember to think about DNA in this context, but also specifically study...plasmids containing these genes. As with the PIs above, many of these genes were discussed at the 2018 ...chromatin This is not the first Addgene blog post about chromatin--multiple Addgene colleagues and guest...

... scientific communication ecosystem Figure 2: The scientific ecosystem, enriched by open access..., social media, coffee chats with potential collaborators, blogging and more. These each have different...key routes for researchers to share information about the emerging COVID-19 pandemic quickly and openly...awareness of the benefits of preprints. Some worry about faulty work being posted as preprints, assuming ...submission. Preprints at Addgene Preprints are all about sharing scientific knowledge faster and more openly...

...state-of-the-art methods of sex and gender analysis and 2) provides case studies with concrete examples of how.... Gendered Innovations is an international, collaborative project—funded by the European Commission, the...research.  In addition to the studies referenced above, another excellent resource for research design ... whole human population. To learn a little more about where differences in effects on the sexes come from...cost. Especially in animal research, they worry about needing more study subjects and control groups. ...

...glycerol stock to each of 2 96-well microplates (identical copies, see image above). We then select forward...One of the best things about sharing plasmids through Addgene is that we provide an added level of confidence...bp of clean sequence from a typical reaction, or about 250 amino acids worth, so make sure the feature ...specifically to target this region, as outlined above. Avoid primers that bind within or must read through...

...available in the adenovirus section below!   Fig. 2 - Viral vector components of AAV Adenovirus Adenoviruses...aren’t reflected in the common components outlined above for gamma-retroviruses. While not required for lentiviral...and helper plasmids for all the viruses discussed above (gamma-retrovirus, lentivirus, adenovirus, and adeno-associated.... Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 1997. Principles of Retroviral Vector ...

...Era of Personalized Medicine. Genes Dis. 2017 Jun;4(2):43-63. DOI: 10.1016/j.gendis.2017.04.001 Maetzig...clinical settings. AAV is also a cool tool in the laboratory! It is ideal for whole animal administration,... be helpful to you. Interested in learning more about a specific virus? We have you covered with guides..., adenovirus, and AAV. If you’re wondering more about what applications these viruses are good for, we...

...IgM, IgD, and IgG are deleted.     Figure 2: Isotype switching requires DNA recombination of the...blog Learn about the advantages and disadvantages of using monoclonal antibodies Read about other techniques... subclasses: IgA1 and IgA2.  You can learn more about isotype classes in our Antibodies 101: What is an...

... the transcriptional level.     Fig. 2 Tetracycline on/off promoters and modes of action ...which can be used with this approach - read more about them on our CRISPRa blog! Fig. 1 CRISPR ... is a lot of overlap with the systems discussed above, just with more elements of control. Tet promoters...to be site-specifically knocked in. This can be labor intensive and time consuming. If your native protein...

...PMID: 20543841. PubMed Central PMCID: PMC2920597. 2. Nathans, Daniel. "Puromycin inhibition of protein... FUNCAT Professor Schuman’s technique applies metabolic labeling using a noncanonical amino acid, azidohomoalanine...both quantitative and spatio-temporal information about the turnover of specific endogenous proteins in ... interested in neurobiology.  She is passionate about communicating science to all audiences. Right now...

...cellular responses in addition to antibody response 2) very high immunogenicity 3) and long-lasting immune...Viruses as Biological Tools blog can help you learn about the different types of viruses and pick one for ...Addgene.org Browse Our Viral Vector Pages Learn About Viral Vector Biosafety Deposit Your Viral Vectors...

... high-priced commercial devices.     Fig. 2: Workflow for open software (left) vs. open hardware... in science brings stronger networking across laboratory and disciplinary boundaries and can allow for... that are otherwise cut off from the process.   About the author: Benedict Diederich received his PhD ...

...closely mimicking the future application. In figure 2 we provide an example of the comparison of the practical... single amino acid. Without further information about properties like photostability or aggregation tendency...Fluorescent Proteins for Multicolor Imaging Learn about Fluorescent Protein Biosensors Additional Resources...

...3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. PubMed. Find the plasmids from this publication ...with limited packaging capacity. Still confused about the various tRNA expression strategies? No worries... Addgene. If you have any questions or thoughts about this technology, leave them in the comments below...

... would release them into the cytosol. Fig. 2: Endogenous sources of duplexed RNA DNA:RNA hybrids...power of dsRNA for a long time, even before we knew about all the diverse in vitro production mechanisms. ...repetitive sequence. All of the R-loops described above play distinct roles in promoting cellular processes...

...higher quenching of mScarlet emission. Figure 2: Ace-mScarlet is a fusion between the voltage-sensitive... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.    ...tracing using the transgenic zebrafish model, ubi:zebrabow. In this system, cells express various fluorescent...

..., N. and Hamilton, D. 1981. PubMed PMID: 6276558. 2. Site-specific DNA recombination in mammalian cells...discussed below. How does it work? As alluded to above, the Cre recombinase catalyzes the site specific...Combining this with some of the loxP methods described above, a genetic modification can be restricted based ...

...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...