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  1. Antibodies 101: ChIP

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    Blog Post
    ...biology techniques. What’s all the fuss? Well, ChIP’s complexities come not from the techniques used, but...immunoprecipitation (ChIP) is an extremely useful technique that provides insight into protein:DNA interactions...
  2. Antibodies 101: Buffers, Storage, and Conjugates

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    Blog Post
    ...excitation by a laser, which is then read by a detector panel in the machine. Fluorophores are light sensitive...different signaling molecule, that all need to be detected in the same assay. This is one of the most complex...signal "spilling over" into another channel's detection range. Though you can compensate for this when...
  3. A Practical Approach to Choosing the B(right)est Fluorescent Protein

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    Blog Post
    ...fluorescent molecule. A higher brightness increases the detected signal and therefore high brightness is an important...brightness that does not fold well or cannot be detected in a microscopy setup has zero practical brightness...excitation wavelength, available emission filters, and detector sensitivity) and the biological system (temperature...
  4. Multiplex Genome Editing with CRISPR-Cpf1

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    Blog Post
    ...Reversing the orientation of the direct repeat protects the (+) stranded lentivirus RNA from Cpf1-mediated... is one of the latest developments in CRISPR technologies.  It’s a simple and effective method with multiple...CRISPR–Cpf1 Asing a Single crRNA Array." Nature Biotechnology 35.1 (2016): 31-34. Pubmed PMID: 27918548. 2...
  5. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

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    Blog Post
    ...CRISPR technology has changed how scientists edit and control genes, but according to the Broad Institute's...activate essentially any gene,” she says. The technical leap from mutating and deactivating a gene or ... require access to next generation sequencing technology. These libraries are lentiviral in nature, ...
  6. Kiran Musunuru on the Newest TALEN Genome-Editing System

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    Blog Post
    ...to essentially any DNA sequence desired. This technology allows you to create a pair of designer proteins...on traditional homologous recombination. That technology had been used for decades to generate knock-out...That’s the revolutionary aspect: TALENs take a technology that was difficult for all but the most experienced...
  7. Antibodies 101: Polyclonal Antibodies

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    Blog Post
    ...experiments that require multiple antibodies that each detect a different antigen. To pre-adsorb the antibodies...section), it may be advantageous if you are trying to detect a protein in low abundance or whose conformational...Overlooked benefits of using polyclonal antibodies. BioTechniques 65:127–136. https://doi.org/10.2144/btn-2018...
  8. Antibodies 101: Selecting the Right Antibody

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    Blog Post
    ... an antibody that is directly conjugated to a detection substrate, such as a fluorophore, or a conjugated...do you know about the vendor and the vendor’s technical support? An antibody from a vendor that you use...frequently, with a responsive and knowledgeable technical support team, is a safer bet than one from a large...
  9. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

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    Blog Post
    ...Biomedical Engineering, 2020. PubMed PMID: 31937940. Detection of neural activity using a red fluorescent genetically...encoded voltage indicators (GEVIs) are proteins that detect and output the voltage changes in neural activity..., has increasingly been recognized for its biotechnological applications including conversion of CO2 into...
  10. CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt

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    Blog Post
    ...it, his lab at the Massachusetts Institute of Technology has been “working on new experimental and theoretical... collision that’s taking place between CRISPR technology and synthetic biology, the tools he’s making ...all heading? Voigt: The top products out of biotechnology today are things like butanol - very simple ...
  11. Rabies and Neuronal Tracing

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    Blog Post
    ... of us learned in biology is that the body’s architecture and plumbing are important. We started with ...oxygenates the body. However, in neuroscience, this architecture is frequently still an open question. The connections...powerful development in neural tracing. Using these technologies in combination with molecular tools (e.g., optogenetic...
  12. Antibodies 101: Choosing the Right Isotype

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    Blog Post
    ...antibodies and found that across suppliers there was a detection bias of IgG2a>IgG2b>IgG1 (Manning 2012).    ... the anti-mouse secondary antibodies used for detection bind to non-target immunoglobulins naturally present.... Chapter 15 Postembedding Immunocytochemical Techniques for Light and Electron Microscopy. In Methods...
  13. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

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    Blog Post
    ...updated Jul 23, 2020 by Jennifer Tsang. CRISPR-Cas technology is constantly evolving. Variants of Cas proteins...be engineered to “armor” therapeutic phages to protect them from pathogenic bacteria’s CRISPR-Cas system... applications: natural brakes for CRISPR-Cas technologies. Nat Methods 17:471–479 . https://doi.org/10.1038...
  14. Mouse Modeling, Part 2: Breeding and Crossing Mice

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    Blog Post
    ...of our mouse modeling blog series, we covered techniques that can be used to introduce genetic modifications...conditional knockin or knockout mice. While these techniques are each useful for introducing one modification...) and using PCR with the appropriate primers to detect your targeting vector or gene of interest. 2. Breed...
  15. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

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    Blog Post
    ...there were unusual patterns of DNA methylation detected in a human disease: cancer (Feinberg & Vogelstein...the original methylated strand is diluted to undetectable amounts: ergo, DNA demethylation! With optimization...repeat and scFv–TET1 catalytic domain fusions. Nat Biotechnol 34, 1060–1065. https://doi.org/10.1038/nbt.3658...
  16. Meet an Addgenie: Insights into Grad School, Career Paths, and More

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    Blog Post
    ... school when she was hired full time as a Lab Technician. “I was so tired of being in school so I just... is not true at all. Racheal Komuhendo, a Lab Technician, dreamed of becoming a doctor. After taking a...circumstance. She was working as a research and technical assistant, when her life changed––she had a daughter...
  17. Plasmids 101: Modular Cloning

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    Blog Post
    ...described above, and not any particular cloning technique. Though rarer, you can find MoClo that relies ...assembled into many combinations. Even with current technologies, it can be extremely hard to predict how a single... if you want to learn more about this cloning technique! Many thanks to Rob Hurt for his help on this ...
  18. CRISPR in the Clinic

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    Blog Post
    ...There can be no doubt that CRISPR/Cas9 technology has been a breakthrough for the genome-editing field... obstacles for future CRISPR therapies CRISPR technology is rapidly evolving, and many of the challenges...taken advantage of what our current delivery technologies are suited for. For example, lipid nanoparticle...
  19. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    ...very easy — our tech transfer team will communicate with your organization’s technology transfer office... ship the samples to Addgene and contact your technology transfer office about the MTA. Pro Tip! Download...
  20. 5 Tips for Troubleshooting Viral Transductions

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    Blog Post
    ...biological tasks, both of which are factors in the technical challenges they can present. Many researchers ...hail Marys that have helped me troubleshoot some technical obstacles when working with virus: 1. Viral vectors... quantification as measured with branched-DNA technology." Journal of clinical microbiology 37.6 (1999...
  21. Cpf1 Update: Comparison to Cas9 and NgAgo

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    Blog Post
    ...predicted off-target sites, Cpf1 does not mediate detectable off-target cleavage. Most gRNAs directed low-...specificities of Cpf1 endonucleases in human cells. Nat Biotechnol. 2016 Jun 6. PubMed PMID: 27272384 Find plasmids...-Cas Cpf1 nucleases in human cells." Nature Biotechnology (2016). PubMed PMID: 27347757. Find plasmids...
  22. Plasmids 101: FLEx Vectors

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    Blog Post
    ... Figure 1 (modified from Schnutgen et al. Nat Biotech 2003): Basic FLEx switch will express BFP in the... Figure 2 (modified from Schnutgen et al. Nat Biotech 2003): In the presence of Cre, GFP expression is... able to distribute materials containing FLEx technology. Read the genOway press release for more information...
  23. Plasmids 101: Terminators and PolyA signals

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    Blog Post
    ...degradosome, which contains enzymes that cut RNA not protected by secondary structure. It is thought that poly... tail is important for stability of the mRNA, protection from degradation, and is integral to the nuclear...Controls Gene Expression Learn about New Cloning Techniques Read all of our Plasmids 101 Posts Resources...
  24. Tips for a 1st time AAV user (by a Rookie AAV user)

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    Blog Post
    ...the AAV production process. Either SYBR green technology or a TaqMan primer/probe set can be used and ...sample for the DNAseI digest. Use good sterile technique when handling your AAV plasmid standard and AAV...Baiker, A. (2012). Universal real-time PCR for the detection and quantification of adeno-associated virus serotype...
  25. Viral Vectors 101: Producing Your rAAV

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    Blog Post
    ...Baiker, A. (2012). Universal real-time PCR for the detection and quantification of adeno-associated virus serotype...adeno-associated virus vectors with sensitivity sufficient to detect single infectious events. Human Gene Therapy, 15...addgene.org  AAV Titration by qPCR Using SYBR Green TechnologyAAV Titration by ddPCRAAV Purification by Iodixanol...
  26. PCR Applications for Copy Number Variation Assays

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    Blog Post
    ... on and so forth. This also serves as a light technical control - an RQ value of 1.5 means you have either...chemistry (see image, left, of droplet immersion technique, courtesy of CAR15.) Each droplet contains a primer...Molecular Biology Reference Molecular Cloning Techniques References Bell AD, Usher CL, McCarroll SA. ...
  27. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

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    Blog Post
    ...plasmid, using the fragment exchange (FX) cloning technique which uses Type IIS restriction sites (Geertsma...protein of interest. The Seeger lab used the ELISA technique described in their protocol because it is less...protein is added and after a few washes, binding is detected using a colorimetric reaction. After the ELISA...
  28. Viral Vectors 101: Voltage Indicators

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    Blog Post
    ... where rhodopsin functions as both the voltage detector and fluorescent signaler. The pro of this system...several targeting domains. The quencher and voltage detector in this system was dipicrylamine, a lipophilic...high-speed, bright, red fluorescent voltage sensor to detect neural activity. Sci Rep 9, 15878 (2019). doi: ...
  29. Academic vs. Industry Postdocs

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    Blog Post
    ...understanding of the fast, deadline-driven pace of biotech. In other words, industry postdocs already have...Biogen, Amgen, New England Biolabs, Regeneron, and Genentech, among others, and the list is growing. Some post... a position open up. Other companies, such as Genentech, will not. The point is that each company has ...
  30. Cloning Mammalian Cells with the Agarose Method

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    Blog Post
    ...Advantages of the agarose cloning method The agarose technique should be useful to scientists who wish to rapidly... from a population of transfected clones. The technique is applicable to a wide variety of cell lines,...physiological properties. An advantage of this technique is the potentially large number of single clones...
  31. Truncated gRNAs for Regulating Gene Expression

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    Blog Post
    ...Biologically Inspired Engineering. CRISPR/Cas9 technology has revolutionized the fields of molecular biology...discovery through the creation of innovative technologies. To date his group has develop several popular...catalytically active Cas9 nuclease." Nature biotechnology (2015). PubMed PMID: 26436575. PubMed Central...
  32. CRISPR 101: Mammalian Expression Systems and Delivery Methods

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    Blog Post
    ...updated Sep 30, 2020 by Marcy Patrick. CRISPR technology has been widely adopted for genome editing purposes...cheaper, faster, and easier than prior editing techniques. More and more CRISPR tools are being published...how you will evaluate the outcome: Will you be detecting insertions/deletions using a mismatch repair assay...
  33. Simplify Cloning with in vivo Assembly

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    Blog Post
    ...experimental process. The majority of current cloning techniques involve the assembly of a circular plasmid in... 1991, Bubeck et al., 1993). But why did this technique fail to gain traction? It is likely that both ...ends on DNA using polymerase chain reaction." Biotechniques 10.1 (1991): 62-66. PubMed PMID: 2003926. 2....
  34. AAVs in Retinal Gene Therapy

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    Blog Post
    ...therapy. The year was 2001 and I started a job as a technician in a lab using adeno-associated viral vectors... I quickly became fascinated by this emerging technology and its potential for the treatment of some genetic...research is being done to further improve the technology: engineering of new capsid variants, designing...
  35. A Guide to Starting Your Own Journal Club

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    Blog Post
    ...club occurs with a small technical team (single lab or team), a deep technical dive on single papers could...must stay current with literature and emerging technologies to remain competitive and relevant. Enter: the...
  36. AAVs CREATed for Gene Delivery to the CNS and PNS

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    Blog Post
    ...CNS and PNS.That is until the Gradinaru Lab at Caltech stepped up to the challenge. Find the PHP Plasmids...Recombinase-based AAV Targeted Evolution. In this technique, a pool of mutant AAV capsid (cap) expressing ...widespread gene transfer to the adult brain. Nature biotechnology. 2016;34(2):204-209. PubMed PMID: 26829320. ...
  37. Plasmids 101: Colony PCR

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    Blog Post
    ...key steps to colony PCR are: 1) design primers to detect the presence of your insert; 2) set up a standard...Running a no template control PCR reaction for detecting contamination is also a good idea.    Analyzing....org Read about Choosing a Molecular Cloning Technique Learn to Perform a Bacterial Transformation Deposit...
  38. CRISPR Antimicrobials

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    Blog Post
    ...reduce this likelihood. The key problem with these techniques is efficient delivery, as the antimicrobial must...efficiently-delivered RNA-guided nucleases.” Nature Biotechnol. 32(11): 1141-5. (2014). PubMed: PMID 252409288...produce sequence-specific antimicrobials.” Nature Biotechnol. 32(11): 1146-50. (2014). PubMed: PMID 25282355...
Showing: 401 - 440 of 795 results