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We narrowed to 823 results for: lars

Showing: 201 - 220 of 823 results
  1. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ...including the popular BL21(DE3) (Li et al., 2021). This pEcCas/pEcgRNA system works similar to the original...modifications, efficiency can be low, especially for larger modifications. What’s the solution to this inefficiency...recombinogenic than E. coli, so this system functions more similarly to CRISPR/Cas9 systems adapted for eukaryotes ...promoters are ubiquitous, they are not universal, particularly when it comes to non-model bacteria. The authors...engineer the bee gut microbiome. They created a modular system using a broad host range plasmid that can... specific locus. This approach provides a more granular level of control where individual bases can be...Bee Gut Microbiome Bacteria with a Toolkit for Modular Assembly of Broad-Host-Range Plasmids. ACS Synth...
  2. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    Published
    Jan. 8, 2019, 1:54 p.m.
    ...reagents with the scientific community.   (click to enlarge) As we look ahead to the things to come, we wanted...total? Just over 49,000 times! Their long-lasting popularity illustrates the utility these tools have in many... The S. pyogenes Cas9 (SpCas9) has been widely popular in genome engineering. For use in human cell lines...plasmid. Indeed the first version is the 15th most popular plasmid of all time, but was found to have a point...publication but its name was subsequently changed to clarify that it does not contain the trademarked Tet-On...lentiGuide-Puro. pSpCas9n(BB)-2A-GFP (PX461) - Similar to pSpCas9(BB)-2A-Puro (PX458), pSpCas9n(BB)-2A-GFP...gRNA. It’s the former version of our sixth most popular plasmid pSpCas9(BB)-2A-Puro (PX459) V2.0. It contains...
  3. Developing a cold tolerant E. coli using specialized chaperones

    Type
    Blog Post
    Published
    Jan. 28, 2020, 2:35 p.m.
    ...face very similar issues when forced to express in mesophilic host systems. The so-called “regular” temperatures...at suboptimal ones, which could allow even your regular E. coli to thrive in the bitter cold.  Since...vs. extremophiles Mesophilic organisms are your regular next-door microbes which grow optimally in moderate...psychrophilic enzymes for milk processing, fruit juice clarification, meat tendering, and oligosaccharide synthesis...the many proteins which are not compatible with regular mesophilic systems are the psychrophilic enzymes...incorporate cold-tolerance inducing genes within a regular mesophilic host system. This would make the host...
  4. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    Published
    Aug. 5, 2025, 1:15 p.m.
    ...heavy machinery. But not all engineering is at a large scale — some is done on much smaller equipment on...depth is antibody directed evolution. This is a larger process of gradually changing and selecting the...Degenerate primers Using degenerate primers is similar to error-prone PCR. Degenerate primers are a group...generation. To evaluate improved properties across large mutant libraries, high-throughput screening techniques... Innovation (IPI) provides information on their large-scale recombinant antibody production platform. ...screening, and affinity maturation. Methods in Molecular Biology, 29–62. https://doi.org/10.1007/978-1-...conformationally selective nanobodies. Nature Structural & Molecular Biology, 25(3), 289–296. https://doi.org/10.1038...
  5. A Guide to Getting Started in Undergrad Research

    Type
    Blog Post
    Published
    June 8, 2021, 1:15 p.m.
    ...duties Staff scientist: similar role to a postdoc but more permanent Visiting scholar/student: visiting from...conferences. The Barry Goldwater scholarship is also a national scholarship that supports students interested...because they had close relatives suffering from similar diseases. Others were inspired by the imminent ...that you would be interested in can also help you clarify your goals and define next steps! More than serving...online for a while and observed. Apply for those scholarships that you don’t think you’re qualified for. Wayne...populations. Research universities often have scholarships and funds dedicated for undergraduates interested...
  6. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    Published
    Sept. 24, 2019, 12:55 p.m.
    ...difficulties in amplifying a template, particularly from a large genome, you can recheck your primer design...plasmid cloning Read our molecular biology reference Find how-to videos for molecular biology and lab techniques...sequence, or if you want to amplify sequences from a large genome organisms like mouse or human. There are ...nanodrop to assemble the  fragments based on their molar ratio. Assembly of the fragments There are multiple...the backbone plasmid; a starting point could be a molar ratio of 1:2 (plasmid:insert), but this parameter...
  7. Antibodies 101: Selecting the Right Antibody

    Type
    Blog Post
    Published
    March 15, 2022, 1:15 p.m.
    ...Antibody Validation proposed 5 pillars for antibody validation.  The 5 pillars include genetic strategies ... project aimed at understanding how a critical cellular pathway is regulated. You’ve read all the background...successfully used in conditions that are the most similar to yours. Take a look at the publications cited...application, a publication successfully used it in a similar way as you plan to. To secondary or not to secondary...technical support team, is a safer bet than one from a large distributor that may not have direct experience ...been validated using one or more of the proposed pillars.  Remember, validation does not mean that the antibody...
  8. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    Published
    May 30, 2023, 1:15 p.m.
    ...experiments ranging from transient gene expression in a particular cell type to CRISPR genome editing, optogenetics... also be used to ensure specificity; this is particularly helpful when wanting to target specific neuron...vector. Full-length ssAAV vector genome of both polarities are rescued from the vector plasmid and individually... conversion of the DNA goes through the inter-molecular annealing or second strand synthesis. In contrast...transcriptionally active double-stranded form through intra-molecular annealing. Figure and legend from Okada, 2013....Production System: Towards Gene Therapy For Duchenne Muscular Dystrophy. InTech. doi: 10.5772/53023 Veldwijk...method for the titration of AAV-2 vector stocks."Molecular Therapy 6.2 (2002): 272. PubMed PMID: 12349826...
  9. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    Published
    June 8, 2017, 2:30 p.m.
    ...Goedhart and Marieke Mastop from the Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced... can be excited many times, thereby producing a large number of emitted photons before they are destroyed...absorb the excitation light less efficiently. Similarly, decreasing the power of the excitation light ...these factors, environmental conditions such as cellular redox state and oxygen concentration may affect...Goedhart is an assistant professor at the Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced...Marieke Mastop is a PhD candidate at the Section of Molecular Cytology (University of Amsterdam) where she develops...Backbones with Fluorescent Tags Find Plasmids for Subcellular Localization Browse the entire Fluorescent Protein...
  10. Plasmids 101: Cre-lox

    Type
    Blog Post
    Published
    Jan. 13, 2015, 3:47 p.m.
    ...two are present; intramolecular events happen with greater frequency than intermolecular events, but any... kDa protein responsible for intra- and inter-molecular recombination at the loxP recognition sites. A...sequence between the loxP sites is excised as a circular piece of DNA (and is not maintained).  Translocation... is excised, and gene expression proceeds. One popular lox-stop-lox plasmid is from Tyler Jack's lab: ...purposes, including activating an oncogene only in a particular organ, or bypassing embryonic lethality. To ...site-directed recombination technology very conceptually similar to Cre-lox, with flippase (Flp) and the short flippase...
  11. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    Published
    April 16, 2024, 1:15 p.m.
    ...Common self-labeling tags Let’s take a look at some popular self-labeling tools: Tetracysteine: The first reported...variants run up to HaloTag11. HaloTag is now the most popular self-labeling tag in Addgene’s collection. CLIP-tag...on the protein of interest. Table 1: Summary of popular self-labeling tag systems Tag Size (amino... HaloTag for their "ticker tape" method for intracellular recordings, switching between different colored...Cornish, 2011). HaloTag and SNAP-tag are the most popular self-labeling enzymes in Addgene’s collection, ...that self-labeling protein tags are increasingly popular. Embrace the flexibility and switch things up for.... Two-colour live-cell nanoscale imaging of intracellular targets. Nature Communications, 7(1), Article...
  12. With an Eye Towards the Future, We Look Back at the March for Science

    Type
    Blog Post
    Published
    Oct. 24, 2017, 1:54 p.m.
    ...pitfalls of accepting alternative facts. Because, similar to scientists, the legacy of each government and...scientific consensus and calling for billions of dollars in cuts to important research funding.  So when...sacrifice in April because devaluing science has alarming implications for our public policies—many of which...that the community of science supporters is much larger and stronger that I expected. Jennifer Stancill...Birmingham, AL, USA 33.5207°N 86.8025°W - Jonathan Aguilar Galileo wrote, “now you would not hide behind these... at the same time pass over in silence his very large one” (http://oll.libertyfund.org/titles/galilei-...deserve to know the truth about our world. Jonathan Aguilar (@AggleDetector on Twitter) is a graduate student...
  13. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ... inherited by daughter cells during cellular reproduction, similar to the host’s chromosome, or they can...both Archaea and Eukaryota can also harbor these circular DNA passengers. Most natural plasmids encode beneficial...transfer can occur naturally through transformation (cellular uptake from the environment), transduction (viral... in a minute! Fun fact! Electroporation is one popular method scientists use to promote transformation...encoding these functions form the basis of many molecular biology tools! Figure 1: Common functions... Mother Nature’s toolbox for new and exciting molecular technologies!  This post was originally written...
  14. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    Published
    Aug. 20, 2019, 12:31 p.m.
    ...to be large enough to target larger areas, typically a few hundred nl. Alternatively, for larger brain...activity, commonly via light-activated ion channels. Similarly, chemogenetic tools are designed channels that...also reduce expression levels of certain genes. A popular method uses Cre recombinase on transgenic flox ...lentiviral vectors are superior packaging vectors for larger genes - they can hold up to 8 kb, while AAV can...liquor of the cerebral ventricles (ICV, intracerebroventricular) to minimize tissue damage; however, the...
  15. Lab Automation at Addgene

    Type
    Blog Post
    Published
    March 22, 2018, 1:28 p.m.
    ...and uses the similar buffers as would be used for a mini prep, except it does it on a larger scale. Using...for standard molecular biology techniques Browse our video library Check out our molecular biology reference...able to do high throughput DNA preps and other molecular work if we need it to. This gives us the oppotunity...the Biomek FxP is it’s customizability. It has a large deck that can house many different types of devices... DNA purification We're developing ways to prep large quantities of DNA because some countries just can...
  16. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    Published
    Aug. 30, 2016, 2:30 p.m.
    ...Biodesign Center for Molecular Design and Biomimetics and the School of Molecular Sciences at Arizona ...In response, the World Health Organization has declared a global health emergency and called for the fast-tracked...sensors within an extensive infrastructure of molecular and electronic companion technologies that has...1x106-fold lower than before), and our combined biomolecular tools can distinguish between global strains...detect the virus in a clinically relevant setting. Molecular diagnostics for the Zika virus predominantly look...Low-Cost Detection of Zika Virus Using Programmable Biomolecular Components." Cell (2016). PubMed PMID: 27160350...
  17. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    Published
    July 23, 2015, 2:30 p.m.
    ...coding the molecular biology of the worm into a computer model that recapitulates how molecular biology ...get involved a in variety of aspects from the molecular biology to learning how to grow and care for the...but we need to develop the manual skills at the regular microscope, and the scripting skills at the automated...locomotion. Our most recent models of C. elegans molecular biology are based on the standard worm genome....Center (CGC) at the University of Minnesota has declared our makerspace an educational institution meaning...Silicon Valley as a student and then instructor at Singularity University at NASA Ames, and an instructor at...
  18. Technical Design of a Western Blot

    Type
    Blog Post
    Published
    Aug. 22, 2024, 1:15 p.m.
    ...250 kDa Similar in size 8% 20–250 kDa Similar in size 10% 10–100 kDa Similar in size ...Conversely, if your protein of interest is large, look for a larger pore size (0.45 µm).  Types of transfers...important part of your western is your antibodies, particularly your primary antibodies. We always recommend...When choosing your lysis buffer, consider the subcellular location of your protein. Proteins located in...to overnight to run, and is recommended for very large proteins. A semi-dry transfer is faster — 3–30 minutes...differing amounts of available epitopes for that particular antibody, this truly does provide a relative,...
  19. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    Published
    April 26, 2022, 1:15 p.m.
    ...noise, due to the large amount of dsDNA in most starting materials, and is not particularly sensitive. The...Protocol: How to Design a primer  Molecular Biology Reference Molecular Cloning Techniques References Bell... copy number assays for years. qPCR works like regular PCR, just with signaling molecules added to the...be affected by DNA quality and primer design, particularly for amplicons under 100 bp (Ma, 2015.) A schematic...ll have to consider:  Resources needed Most molecular biology labs and departments will have qPCR machines...
  20. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ...columns had similar DNA yields. Additionally, columns regenerated up to five times had similar DNA yields...Addgene.org Find protocols for molecular biology and cloning Read our Molecular Biology Reference Visit our...plasmid miniprep protocol from Addgene follows a similar workflow as a column-based plasmid extraction kit... This method provides DNA yield and quality similar to that obtained with commercial kits, but in less...cellulose-based method was faster, cheaper, and had a similar level of sensitivity. Figure 2: Steps to ...
Showing: 201 - 220 of 823 results