Skip to main content
Addgene
Open Menu

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

Information

In Category

We narrowed to 629 results for: EMB

Showing: 101 - 120 of 629 results

  1. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ... would be injected into developing mouse embryos, the embryos allowed to develop into chimeric mice (with...would first introduce a mutation into a mouse embryonic stem cell (ESC) line, enrich and select for cells...fascinated by CRISPR and rushed to test it. I still remember the day, March 16, 2014, that I got my first preliminary...Streptococcus aureus, is as effective at editing mouse embryos as Cas9 from S. pyogenes and has the advantage ...of fertilized mouse eggs. Or, to avoid handling embryos ex vivo, the components can be electroporated into...Doench’s gRNA design blog post. Finally, always remember that you are working with RNAs, which are prone...payload! Last but not least, when using CRISPR, remember to appreciate the fact that it was first discovered...

  2. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    .... They can be targeted to the plasma membrane to measure membrane potential and can also be directed to... the membrane), the voltage sensing component (i.e., rhodopsin naturally resides in the membrane), or ... want to monitor the voltage change across the membrane of cardiac cells. Similarly, neuronal signaling...sensors contained a FP that localized to the plasma membrane due to the addition of several targeting domains...compound that distributes itself within the plasma membrane as well. Another flavor of these tools is Voltron...more lipophilic, increasing its affinity for the membrane, where it will then localize and emit signal from...generate without disturbing the integrity of the membrane, a concern with several of the previously described...

  3. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    ...contacts that bring apposing membranes together and can wrap the membranes into stacked structures including...dimerization.   1. Transmembrane fluorescent protein fusions Transmembrane proteins, i.e. receptors...they cannot tumble in 720°, i.e. they are on a membrane or in an organelle, then their effective concentration...dimerizing fluorescent proteins is illustrated. 1) Membrane proteins fused to FPs that dimerize can form dynamic...integrated into lipid bilayers. Fusion of an FP to a transmembrane protein significantly increases the effective...other. The consequences can be dramatic. For a membrane POI localized to the endoplasmic reticulum, the...tubules can be grossly distorted into dense stacked membranes termed Organized Smooth Endoplasmic Reticulum ...

  4. Cultivating Community Science at BosLab

    Type
    Blog Post
    ...twenty members and volunteers. Transforming citizens into scientists Anyone can become a member of BosLab... if we were teaching classes at the lab, so in December 2015 we held our first intro to bacterial transformation...previous laboratory experience is not required. Our members are skilled with audiovisual media, software engineering...instructor for our intro to biohacking courses. Our members pursue a diverse range of projects at BosLab. Some...experience targeting RNA with small molecules. Other members’ projects range from hacking probiotics to filming.... Community projects at BosLab Not all of our members start out with personal projects in mind. Thus,...encourage more involvement and provide all our members the opportunity to do molecular biology, we’ve ...

  5. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    ...appropriate vector. Gibson Assembly, GeneArt Cloning, and SLIC can all be used to assemble multiple fragments,...insertions require complicated, often costly in vitro assembly methods. A new method, REPLACR-mutagenesis, harnesses...substitutions - at the same efficiency as Gibson Assembly and GeneArt cloning - but at a much lower cost...mutagenesis methods Current SDM methods rely on in vitro assembly. To make a point mutation, you design two oligonucleotide...al. wondered if, instead of relying on in vitro assembly, they could make the E. coli do the work by using...expensive than gBlock synthesis, and multifragment assembly methods are preferred. No such restriction applies...REPLACR-mutagenesis goes head-to-head with Gibson assembly and GeneArt cloning, the results are favorable...

  6. Gaining Leadership Skills Volunteering at a Professional Organization

    Type
    Blog Post
    ...laboratory work). Committee members have to work professionally with the other members on the team and keep ... according to the needs of my committee members. Some members would need much more guidance, direction...meeting. I first joined a committee as a volunteer member. The NPA runs its committees through Chairs and...and Vice-Chairs that lead them. As a committee member, I volunteered for various tasks as things came up...Directors and liaise with the Executive Director. As members changed, I had to learn how to develop new working...particular style and requirements of the new Board Members. In some instances I felt like I was going back... speaks in “one voice” whilst among individual members, many opinions may exist. I learnt to try and bring...

  7. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    ...successfully modified zygotes or embryonic stem cells. Modified embryonic stem cells are injected into a...techniques used to create engineered mouse models at embryonic stages, and the pros and cons of different genome...containing the gene of interest into a zygote, or into embryonic stem cells that are then injected into a blastocyst...insert the gene of interest into a zygote or into embryonic stem cells. Non-homologous recombination incorporates...recombination. A targeting vector is electroporated into embryonic stem cells and this will insert the gene at a ...mice, knockout mice are engineered during the embryonic stem cell stage using one of several techniques...technique is the injection of a targeting vector into embryonic stem cells, which are then injected into a blastocyst...

  8. Networking and Inspiring Young Scientists at Freshers Fairs

    Type
    Blog Post
    ...has 1,283 members, who comprise undergraduates, graduate students, postdocs, faculty members, and the ...rides and other fun activities and even invite members of the public to attend. We thought we’d take some...study chemistry. The society currently has 352 members, ranging from undergrad all the way up to professors... University of Oxford currently has around 700 members and hosts a range of scientific meetings for students...also hosts high level academic debates for its members to become involved with. The society provides talks...in this emerging field. It has gained over 170 members in its first year. The society facilitates opportunities...education and networking opportunities for society members as well as arranging collaborations and conferences...

  9. Plasmids 101: Gram Negative and Positive Bacteria

    Type
    Blog Post
    ...diderms, have a thinner peptidoglycan membrane and an outer lipid membrane. Common gram negative bacteria include...within their membrane, while the stain is washed from the gram negative bacteria’s thinner membrane. Procedure...destroys the outer lipid membrane, and the thin inner peptidoglycan membrane is unable to retain the stain...or monoderms, have a single thick peptidoglycan membrane. Common gram positive bacteria include staphylococci...interaction of CV+ and I- in the thick multilayered membrane of the gram positive bacteria. In a gram negative...

  10. The Crispr Babies Saga Shows the Need for Action, Not More Delays

    Type
    Blog Post
    ...edit a human embryo, or have a woman carry an edited embryo to term. (We should remember that the woman...these embryos is actively being experimented on, too.) However, as research published on December 13 shows..., founding bioethicist of Rogue Bioethics. On November 25, the MIT Technology Review dropped a bombshell...techniques that are sufficiently safe to manipulate an embryo—even if it’s not necessarily via CRISPR/Cas9. This...unethically experiment on humans, and they are not remembered kindly. However, despite many headlines questioning...

  11. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    ...consists of modular parts that are assembled with Golden Gate Assembly. They built this toolkit upon the...vast, the ability to genetically alter microbiome members is limited and many microbes are difficult to culture...method for genetically altering gut microbiome members in their native habitat (Ronda et al., 2019). They...microbiome systems. Find the toolkit for modular assembly of broad host range plasmids Counting cell division...distributed cell division counting (DCDC), relies on the assembly of fluorescent proteins into one bright particle...fused the red fluorescent protein (RFP) to a self-assembling protein (SAP) such as a bacteriophage shell protein...microbiome bacteria with a toolkit for modular assembly of broad-host-range plasmids." ACS synthetic biology...

  12. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    ...allowing you to easily create customized overhangs to assemble up to 6 different fragments in a prescribed order...few dollars per reaction. By comparison, Gibson assembly can cost over $15.00 per reaction if you purchase...purchase a pre-made mix. Nonetheless, Gibson assembly is a highly versatile and efficient way to generate...such as cDNA, PCR fragments, or genomic DNA. Also remember that commercially available cloning kits can often...feature biases the reaction in favor of the desired assembly to make the process close to 100% efficient. Gateway...declines as the number of fragments you’re hoping to assemble increases. Multiple inserts: There are some cloning...through sequence homology. Similarly, Golden Gate assembly allows one to simultaneously complete an ordered...

  13. Congratulations, Deck The Lab winners!

    Type
    Blog Post
    ...their lab’s decoration competition, and each lab member used an impressive variety of lab materials to ...drNBUdlQco — The Tarling-Vallim Lab (@TarlingVallim) December 19, 2020 In third place, the BTC Institute decorated...pic.twitter.com/OOrwo9feyJ — BTC Institute (@BTCInstitute1) December 23, 2020   Deck the Lab: Home Edition Winners...safe and festive Holiday season. 🙌 🥂 🎄 🎉 😷 Remember wear a mask,↔️social distance, 💓show gratitude...pic.twitter.com/ofLI4eD8Zs — Microbigals (@MicrobiGals) December 23, 2020   In second place, Dr. Anne Madden created.../GsAlD6sjM6 — Dr. Anne A Madden (@AnneAMadden) December 23, 2020   In third place is Matthijs Dorst’s...pic.twitter.com/qDylF8TdHH — Matthijs Dorst (@MattNeuro) December 4, 2020 Congratulations to the winners! We’ll...

  15. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...rate limits its use in systems like the C. elegans embryo, where developmental processes happen within a ...expression levels comparable to GFP during early embryogenesis (Figure 3). mScarlet-I3 also offered clearer...fluorophore-tagged GOLG-4 through eight stages of embryonic development. Representative images of single focal... focal planes through the middle of each embryo are shown, comparing GFP, wrmScarlet (mScarlet), and the...the 11-amino acid BiT fragment of luciferase is embedded in the I10 domain of the human titin protein and...LgBiT luciferase fragment (expressed separately) assemble and catalyze the luminescence reaction (Figure...of luciferase binds to the BiT Tag (also cyan) embedded in the Titin I10 protein (purple) only when Titin...

  16. Management for Scientists: Seeking Feedback

    Type
    Blog Post
    ...excellent, effective communication between all members of a team. As described here in a 2012 Intuit blog...conducts exceptionally well-managed meetings. Members face one another, and their conversations and gestures...well as give, information to make improvements. Members connect directly with one another – not just with...be covered in a future post in this series). Members carry on back-channel or side conversations within...everyone knows what is going on and can contribute. Members periodically break, go exploring outside the team...that brings feedback back to the manager. Team members rarely rush to give honest feedback to the manager...over coffee.  Have regular 1:1 meetings with team members (don’t become known for cancelling these); ask ...

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

    Type
    Blog Post
    ...achievement Evidence of membership in associations that require their members to demonstrate outstanding...with US immigration in its current state. As of September 2017, USCIS received 116,224 employment based ... position. If you cannot get an Industry H-1B, remember that industry career paths are not the only place...the lab or has won, awards, media reports, and membership in groups related to the foreign national’s field... national award for excellence in your field. Membership in a professional organization – this is not ...assist your PI in the peer review process, but remember you cannot use it as documentary evidence for ...point of view of experts in your field. Please remember, this is not a job reference letter. This letter...

  19. Synthetic Biology - Algal

    Type
    Collection
    ...SynBio Resources SynBio Home Browse All SynBio Assembly Standards SynBio Blog Genome Engineering Browse...

  20. Synthetic Biology - Bacterial

    Type
    Collection
    ...SynBio Resources SynBio Home Browse All SynBio Assembly Standards SynBio Blog Genome Engineering Browse...
Showing: 101 - 120 of 629 results