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We narrowed to 604 results for: abo.2

Showing: 321 - 360 of 604 results

  1. Pooled Library Amplifications

    Type
    Blog Post
    ...estimate transformation efficiency (Figure 2).    Figure 2: Example of a serial dilution to estimate...First, you’ll need to collect specific information about your library: the size of your library (How many...with Addgene. If you have any specific questions about designing your library amplification protocol, feel...

  2. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ...mean you have one copy of the gene; an RQ value of 2 means you have two copies, and so on and so forth....quantitative PCR. Expert Rev Mol Diagn. 2005 Mar;5(2):209-19. doi: 10.1586/14737159.5.2.209. PMID: 15833050...controls must be run in triplicate.  The nice thing about qPCR is that since it measures relative quantification...number, albeit within a range of RQ values as noted above.   In both applications, results can be affected...

  3. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ... of permeabilization and membrane integrity (Fig. 2). For example, saponin interacts with cholesterol ... at too high a concentration.      Figure 2: Examples of different permeabilization strategies...and detergents.  Organic Solvents  As mentioned above, organic solvents can be used to simultaneously ...permeabilizing with saponin. However, if you are not worried about washing away your target (for example, some cytoskeletal...

  4. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...from GFP and the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising..., V163A EBFP F64L, S65T, Y66H, Y145F   Table 2: Functional role of specific mutations in GFP derivatives... activity, and much more. Read on to learn more about GFP, how scientists have evolved this versatile ...effects of alterations to this activity. Learn more about optogenetics at OpenOptogenetics and find optogeneic...

  5. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...gamma-retroviral vectors, you need three plasmids (Figure 2): Transfer plasmid — contains transgene, sgRNA, or...for different cell types (known as tropism). Figure 2: Gamma-retroviral plasmids. Created with BioRender.com...Read our gamma-retroviral guide to learn about gamma-retroviral components, retroviral production, and...gamma-retroviral vectors that can be safely used in laboratory settings. These engineered gamma-retroviral vectors...of plasmids, as lentiviral vectors do. For more about lentiviruses, see Addgene's lentiviral vector guide...increased safety when producing viral vectors in a laboratory setting. Figure 1: Wild-type gamma-retrovirus...packaging methods . The three plasmids described above (envelope, packaging, and transfer) are co-transfected...

  7. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...Indicators GEVIs can be divided up structurally into 2 main groups: voltage sensitive domains and Rhodopsin...depending on your experimental system.   Figure 2: General overview of chemigenetic tools. A genetically...especially essential. If you want to learn more about how viruses can be used for chemigenetics, biosensors...

  8. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    ...of ~75% in response to the magnetic field (Figure 2); they named this variant MagLOV. Andrew was thrilled...didn’t need any added cofactors.      Figure 2: Fluorescence intensity and ΔF/F of MagLOV, a variant...our tissues unimpeded. Andrew and Maria dreamed about the possibilities of a magnetoresponsive fluorescent...of improved response. Then, just when Andrew was about to suggest she deprioritize the project, Maria’s...with a strong commitment to exploration and collaboration. Andrew is a physicist with a background in ...potential applications.  Andrew is a frequent collaborator and is happy to give advice and support for ...interested in working on these questions or collaborating, the York Lab is eager to hear from you. Find...

  9. New Podcast Segment: Hot Plasmids

    Type
    Blog Post
    ...mutations. Listen to Our Segment on CRISPR X Hot Plasmid 2: Magneto 2.0 form Ali Guler's Lab - Control neuronal...this new way to learn about plasmid technologies. If you have any comments about the Hot Plasmids Podcast...with these efforts, you’ll find new ways to learn about science careers, lab protocols, and, of course, ...each one, plug in some headphones to learn more about the latest and greatest plasmids through this podcast...

  10. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...with neuron-specific expression of dCas9-VPR (Figure 2), a popular Cas9 activator developed by the Church...ubiquitous ones such as EF-1α and CAG. Figure 2: Dual lentivirus system with ubiquitous expression... to target individual genes can be relatively laborious because multiple plasmids/viruses are needed to...screening strategies Although the tools described above make CRISPRa more accessible to neuroscientists,...vitro and in vivo applications, the tools described above make CRISPRa in the brain relatively easy and accessible... CNS using CRISPRa as a member of the Gelbard Laboratory at URMC.     References Liu, Yanxia, et al. "...

  11. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ...connectivity to other data sources used in research; (2) connectivity to other tools used in research; and...by making it easy to get data out of the ELN; and (2) facilitate inter-operability with other research ...RSpace is integrated with most of the tools noted above, and has a modern API. Integrations with additional...into RSpace. The kind of informal conversations about research that can take place in Slack normally never...RSpace documents to external data sources (described above) further enhances the value the repository integrations...Minisymposium on Reproducibility Read all blog posts about scientific sharing Resources on Addgene.org Find...

  12. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    ...diffusion test or Kirby-Bauer susceptibility test (Figure 2), and requires at least 18-24 hours to obtain results...." London: Wellcome Trust & HM Government (2016). 2. CDC Information Pages on Antibiotics 3. WHO Antimicrobial...resistant infection or not, because scientists in the laboratory will need to isolate and grow whatever bacteria...method that is frequently used in the clinical laboratory. What can we do to stop the emergence of antibiotic...CDC. Antibiotic resistance can be scary to think about, but we can all do our part to learn all we can,...drug resistance. Additionally, she is passionate about health equity, science communication, and public...

  13. An Interview with Connie Cepko - Gene Therapy, Plasmid Tools and Insights to Success

    Type
    Blog Post
    ... Find plasmids from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “Cell type-specific manipulation... Cepko. Listen to the podcast below to hear all about the research being conducted in Professor Cepko'...and mentoring styles. If you'd like to learn more about Professor Cepko's recent work on developing GFP-...the time to talk to us and look forward to hear about more exciting discoveries from her lab. Special ...

  14. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...green squares, purple circles and blue triangles. 2) An antibody immobilized on a bead binds specifically...protein G, a novel IgG-binding reagent. J Immunol 133(2):969-974. Hjelm H, Hjelm K, Sjöquist J (1972) Protein...proteins or nucleic acids. Read on to learn more about this versatile technique.  Immunoprecipitation ...proteins in an IP are not denatured (learn more about antibodies here!) When using Protein A/G to immobilize...eluted by boiling the beads in SDS. (To learn more about Western blotting check out our blog post The Basics...

  15. Plasmids 101: How to Name Your Plasmid in 3 Easy Steps

    Type
    Blog Post
    ..., visit Vector Database. Step 2: Insert name  Include information about the insert in your plasmid name... just by reading the name. For more information about plasmid features and tips you can use in the lab...

  16. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...antibiotic selection. Essentially, this gene is a 2-in-1 providing both bacterial and eukaryotic selection...Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage/gei016I ...therefore prevent cell death. One of the cool things about amp is that it's exclusively toxic to dividing cells...antibiotic selection. If you feel like multiple of the above antibiotics would work for you, they likely do! ...

  17. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ...expressed as a fusion with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that ...cell 58.4 (2015): 644-659. PubMed PMID: 26000849. 2. Grimm, Jonathan B., et al. "A general method to improve... the frontiers of biological imaging.  *Worried about your budget? Email [email protected] Resources on the Addgene Blog Learn About Other Protein Tags Catch Up on Your Fluorescent ...

  18. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...retain their individual activities when separated (2), and that DBD and TAs from different systems could... Enzymol. 1983;100:293-308. PubMed PMID: 6312261. 2. Keegan, L., Gill, G., and Ptashne, M. Separation ... Plasmids Read our Blog Post About Yeast Vectors Read our Blog Post About Tagging Yeast Genes ...

  19. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ...of these neurons via the channel Channelrhodopsin 2 (ChR2) reduced glutamate transmission from the mPFC...deliver Cre-dependent expression of  Channelrhodopsin 2 (ChR2), a modified receptor protein that activates...distant regions. They also don’t give any clues about the types of neurons involved or whether their involvement... Viral Vectors Resources on Addgene.org Learn about Addgene's viral service Find viral protocols and...

  20. PEI Calculator for Planning AAV Packaging Transfections

    Type
    Blog Post
    ...pHelper: 1 RepCap OR 3 Transfer Plasmid : 5 pHelper: 2 RepCap Select what type of tissue culture dish you...packaging experiments. Let us know what you think about the calculator in the comments section below. References...

  21. Countdown to Halloween @Addgene

    Type
    Blog Post
    ...Halloween is only 2 days away and here at Addgene we couldn’t be more excited! As a small, tight-knit...need more convincing, I’ve got just the proof: Above our office team shows their love of plasmids! Each...

  22. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...linear region to solve for total expression time. 2. Timer functionality with a single gene Although it...290.5496 (2000): 1585-1588. PubMed PMID: 11090358. 2. Tsuboi, Takashi, et al. "Age-dependent preferential...Additional Resources on the Addgene Blog Learn More About Fluorescent Protein Aggregation Catch Up On Your...

  23. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...detection, and control. Int J Pharm. 2018 Oct 25;550(1-2):251-268. doi: 10.1016/j.ijpharm.2018.08.043. Epub...and particle formation. J Pharm Sci. 2015 Feb;104(2):602-11. doi: 10.1002/jps.24259. Epub 2014 Nov 21....validated, it’s ready for use! You can learn more about common antibody applications in our Antibodies 101...

  24. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ...et al., 2020). Genomic targets require an upstream 2-bp PAM sequence for recognition by QCascade; although...cells of the plasmid after integration. Figure 2: (A) Streamlined VchINT expression constructs. (B)..., 2020), to multi-copy genomic integration of metabolic cassettes (Zhang et al. 2020); however, the technology...

  25. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...your sample (i.e. cultured cells or tissues).  Step 2: attach anchors to your sample that will link biomolecules...quite so much. Created with biorender.com Steps 2-4 are not found in standard IHC/ICC protocols and ...applications and there’s just not much to be done about that.  Second, expansion introduces distortions....

  26. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    ...efficient knockout (KO) of genes in immune cells; (2) is not immunogenic, a necessity for in vivo studies...faithfully model the complex spatial, cellular, metabolic, and cytokine microenvironment that exists in ...LaFleur is a Postdoctoral Fellow in Arlene Sharpe’s laboratory at Harvard Medical School and is interested in...

  27. Podcast: Interview with Addgene's Founders

    Type
    Blog Post
    ...Intro Chapter 1 - Who are Addgene's Founders? Chapter 2 - The process of founding Addgene, where the idea ...repository’s future. If you’ve ever been curious about what it takes to start your own nonprofit, this ...Harvard University, where she studied diabetes and metabolism. Melina has worked at Genetics Institute in Cambridge...

  28. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    ... both substantial merit and national importance; (2) The foreign national must be well positioned to advance...outstanding achievements in your specific academic field, (2) that you have three 3 years of research experience..., Casseus Law suggests a mix of experts including 2 dependents –people who have worked with you i.e. your... data and the list goes on and on. As you think about your career (and your future in general), keep in...in this post benefit from the applicant thinking about the process early, keeping good records, and planning...writing, compliance, core facility management, collaboration management, grant administration, clinical operations... in professional publications written by others about the alien's work in the academic field Evidence ...

  29. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    ...Information Science and Technology 60.4 (2009): 858-862. 2. Van Noorden, Richard. "Interdisciplinary research...Librarianship and Scholarly Communication 2.2 (2014): 2. Additional Resources on the Addgene Blog 6 Tools...learn about how researchers are using “big data” to look at Lyme Disease in new ways. Or learn about the...ot­her disciplines, are even less likely to read about our work. We therefore must discover new ways to.... Only on occasion do we then have time to read about research from other disciplines, or from other countries...communication through video     Video abstracts about research, especially published research, have grown...researchers have Facebook pages, few share any details about their research itself. Pinterest is also a favorite...

  30. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...faster when using a dual guide system.   Fig. 2: Size screening for large deletion events.   ... Biol 17: https://doi.org/10.1186/s13059-016-1012-2 Find more CRISPR Resources at Addgene: Read our CRISPRGuide... your edit, it’s important to know a few things about your target cells and the desired edits. First, ...et al., NAR). It works just like TIDE (outlined above), except that it uses a 3rd sequencing trace file...great option, especially if the options outlined above aren’t realistic. Check Out Our Post on Sequencing...higher probability locations. If you’re worried about off-target effects, you can consider using a version...SpCas9-HF1, eSpCas9(1.1), or HypaCas9. Read more about Cas9 options in our blog post here. Dealing with...

  31. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    ...’re delivering their payload to.   Figure 2: Numbers of AAV tools, by category, available from... substrates, light, or engineered ligands (Figure 2). Biosensors, which include calcium and voltage sensors...all, a delivery method. I would be remiss to talk about them without looking at what they’re delivering .... But in the years since, we’ve learned so much about what it takes to make these viral vectors not only...., 2016), video, educational resources, and collaborations, and will continue to do so, in the hopes that...leave a comment; or take our AAV survey. Learn more about GCaMPs and other tools developed through the GENIE...

  32. Synthetic Biology, Artificial Chromosomes, iGEM & More - Interview with Tom Ellis

    Type
    Blog Post
    ...balance in Academia. Will it all pay off? Chapter 2: The Centre for Synthetic Biology and Innovation -...synthetic yeast chromosome. Listen to learn more about Dr. Ellis, synthetic biology, synthetic chromosomes...Additional Resources from the Addgene Blog Learn about the Synthetic Biology Education Nonprofit BioBuilder...BioBuilder Plasmids 101: Golden Gate Cloning Learn About Working at a SynBio Startup in Our Interview with...

  33. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...the chromosome, ending replication.   Figure 2: Prokaryotic chromosomal replication. Adapted from...Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/MMBR.62.2.434-464.1998... negative regulators. The genome of E. coli is about 4.6 million base pairs long and contains a single...copy number and compensating for the additional metabolic cost imposed on the host with a beneficial function.... On the other hand, they also impose a lower metabolic cost on cells and produce lower amounts of protein...

  34. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    ...blood are reprogrammed immediately after collection [2]. PBMCs display unique epigenetic signatures that ...PMID: 27882265. PubMed Central PMCID: PMC5118044. 2. Kim, Y., et al., The Generation of Human Induced ...successfully to derive iPSCs from PBMCs. Most laboratories use Sendai virus to reprogram PBMCs efficiently...establishment of virus-free hiPSCs usually require about 5-10 passages. Furthermore, working with Sendai ...source of somatic cells. This translates to only about 16 hiPSC colonies per 100 ml of starting blood sample...

  35. 500,000 Plasmids Shipped and Counting!

    Type
    Blog Post
    ...at Addgene – 500,000 plasmids shipped! That's a 1/2 million plasmids sent to happy scientists around the...scientists outside of the US now use the repository – about 50% of plasmids are shipped to labs outside of the...distribute their plasmids. The partnerships and collaborations with institutions and tech transfer offices...

  36. Addgene's Website Update

    Type
    Blog Post
    ...prompt you to log in or create an account.  Figure 2: The new deposit materials bar New menu Most common...straight from the new search bar, but the menu just above it can help you quickly and easily navigate to educational...down? You’ll find Blugene, as well as information about Addgene, our mission, links to our deposit, request...

  37. Your Top Requested Plasmid in 2015!

    Type
    Blog Post
    ...PMID: 24336571. PubMed Central PMCID: PMC4089965. 2. Sanjana, Neville E., Ophir Shalem, and Feng Zhang...cells are avaliable for distribution. To learn more about pooled library screening, read our recent blog post...the Addgene Blog Browse Our CRISPR Posts Learn about Using CRISPR to Generate Knockouts Read Some Common...

  38. Inntags: Innovative Protein Epitope Tagging

    Type
    Blog Post
    ...tagging." Nature methods (2015). PubMed PMID:26322837. 2. Stadler, Charlotte, et al. "Immunofluorescence and...takes a lot of time and energy, Martí Aldea and collaborators have created a set of “innocuous tags” (inntags...Pubmed PMID: 23435261. Resources at Addgene Read about other Protein Tags in our Plasmids 101 Series. Get...

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

    Type
    Blog Post
    ...sequence is unique compared to the rest of the genome.  2) The target is present immediately upstream of a Protospacer...utilized allowing for PAM flexibility. Fig. 2: Prime editor engaging target DNA. The reverse transcriptase... 101 Series Learn about the PAM Requirements of Different Cas9 variants Learn about CRISPR Pooled Library...

  40. The Addgene Antibody Data Hub

    Type
    Blog Post
    ...whether the results include knockout data.    Figure 2: Search options in the Antibody Data Hub. Data reports... antibody worked under the conditions described above, and a short description of how the antibody performed...
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