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  1. How-to: 5 Steps to a Great Panel Discussion

    Type
    Blog Post
    Published
    Oct. 22, 2014, 1:57 p.m.
    ...uninteresting, the panel members all say the same thing and one panel member dominates the discussion?... no time for audience questions and an audience member asks a 5 minute question that is really a speech... a fun discussion. People are much better at remembering these things on a relaxed phone call than when...moving and to prevent any one panelist or audience member from taking over. My own personal rule is that ... get things started. Give a couple of audience members a chance to participate. For example, at a recent...nonprofit science organizations, I asked audience members to stand and describe unique nonprofit models that...in the audience. I always try to meet audience members before a panel discussion and often incorporate...
  2. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    Published
    May 14, 2024, 1:15 p.m.
    ...Modular cloning is a popular DNA assembly tool used to build single- and multi-gene constructs for a ...is. MoClo background MoClo is a clever way to assemble several DNA parts in a specific orientation using...are reusable, saving time and money on future assemblies. If you are new to MoClo, give our MoClo 101 ...Kit has a wide array of DNA parts that can be assembled into individual TUs or very large (up to 20!) ...CIDAR offers a one-pot reaction wherein multi-part assembly can occur. It is quite extensive with a supplemental... most intuitive CRISPR MoClo application is to assemble multiple unique sgRNAs into a multi-TU targeting...save time and money on studies that require many assemblies or that compare the effects of different DNA ...
  3. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ...diffuse through the cell membrane freely, while others require a permeabilized membrane to enter cells. Depending...Depending on the application, they can be membrane-permeant or membrane-impermeant, and the cells of interest...viability dyes are membrane-impermeant and can only enter cells with compromised plasma membranes — i.e., dead... The targeted epitopes can be motifs within transmembrane proteins, such as receptors, or post-translational... marker of interest is not located on the cell membrane but inside the cell. Due to their chemical nature...nature, antibodies cannot penetrate the cell membrane; hence, intracellular labeling with antibodies requires...Permeabilization, as the name suggests, perforates the cell membrane to allow for antibodies to enter the cell (Figure...
  4. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    Published
    July 12, 2016, 2:30 p.m.
    ... 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...
  5. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    Published
    Jan. 30, 2024, 2:15 p.m.
    .... 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...
  6. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    Published
    April 19, 2016, 2:30 p.m.
    ...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 ...
  7. Cultivating Community Science at BosLab

    Type
    Blog Post
    Published
    Nov. 2, 2017, 1:35 p.m.
    ...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 ...
  8. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    Published
    Feb. 10, 2016, 3:30 p.m.
    ...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...
  9. Gaining Leadership Skills Volunteering at a Professional Organization

    Type
    Blog Post
    Published
    Aug. 6, 2015, 2:30 p.m.
    ...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...
  10. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    Published
    July 11, 2019, 1:26 p.m.
    ...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...
  11. Networking and Inspiring Young Scientists at Freshers Fairs

    Type
    Blog Post
    Published
    Nov. 17, 2015, 3:30 p.m.
    ...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...
  12. Plasmids 101: Gram Negative and Positive Bacteria

    Type
    Blog Post
    Published
    Aug. 2, 2022, 1:15 p.m.
    ...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...
  13. The Crispr Babies Saga Shows the Need for Action, Not More Delays

    Type
    Blog Post
    Published
    Dec. 20, 2018, 1:15 p.m.
    ...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...
  14. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    Published
    June 27, 2019, 12:25 p.m.
    ...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...
  15. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    Published
    Aug. 21, 2018, 12:31 p.m.
    ...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...
  16. Congratulations, Deck The Lab winners!

    Type
    Blog Post
    Published
    Jan. 14, 2021, 1:30 p.m.
    ...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...
  17. Hot Plasmids: Fall 2024

    Type
    Blog Post
    Published
    Nov. 21, 2024, 1:30 p.m.
    ...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...
  18. Management for Scientists: Seeking Feedback

    Type
    Blog Post
    Published
    Jan. 8, 2015, 2:34 p.m.
    ...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 ...
  19. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    Published
    Dec. 20, 2017, 2:33 p.m.
    ...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...
  20. From Lab Bench to Leadership: Transitioning into Management as a PhD

    Type
    Blog Post
    Published
    Jan. 21, 2025, 2:15 p.m.
    ...requires deliberate preparation and a willingness to embrace new responsibilities. After my second postdoc,...differencesManaging a team isn’t “one size fits all”. Each team member has unique motivations, strengths, and learning...scientist, I worked with a diverse team where some members thrived on independence, while others required ... stay on track and feel supported. Another team member was highly self-motivated and excelled when given...to the communication style of a new, junior team member who had values, working styles, and skills that...discovery project, I noticed tension between two team members with differing approaches—one detail-focused and...permission to explore, reflect on your priorities, and embrace these transitions as an integral part of your broader...
  21. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    Published
    April 16, 2024, 1:15 p.m.
    ... with SiR-CA (magenta) and outer mitochondrial membrane labeled with SNAP-OMP25 with Atto590-BG (green... 3: Single-molecule tracking of actin filament assembly-regulating proteins in vitro reveals complex events... probes are not very good at crossing the cell membrane. So, you should investigate your options and weigh...self-labeling protein tags are increasingly popular. Embrace the flexibility and switch things up for your next...Biology, 26(4), 584-592.e6. https://doi.org/10.1016/j.chembiol.2019.01.003 Gallagher, S. S., Sable, J. E., Sheetz...Biology, 15(2), 128–136. https://doi.org/10.1016/j.chembiol.2008.01.007 Griffin, B. A., Adams, S. R., & Tsien...https://doi.org/10.1038/nbt765 Lavis, L. (2016, September 8). Better Dyeing Through Chemistry & Small Molecule...
  22. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    Published
    Sept. 24, 2019, 12:55 p.m.
    ...Gibson Assembly, Golden Gate etc.), and try to find the simplest way to do it (i.e. avoid assembling too ...gel or by nanodrop to assemble the  fragments based on their molar ratio. Assembly of the fragments There...can generate the parts of DNA that you want to assemble together in a combination of different ways according...have a successful and ultimately faster cloning. Remember to quantify the DNA concentration of all the samples...There are multiple ways you can assemble the different parts of a plasmid based on the cloning strategy...
  23. It’s Bioelectric! An Exciting Interdisciplinary Field of Research

    Type
    Blog Post
    Published
    Oct. 29, 2019, 2:52 p.m.
    ... Pai, Vaibhav P., et al. "Transmembrane voltage potential controls embryonic eye patterning in Xenopus...slightly slower scale.  All living cells with a membrane possess an electrical state. This state, known...is maintained by ion channels and pumps in the membrane. An emerging interdisciplinary field of biology... depolarized (positive) the cell is, the more embryonic, stem cell-like, or cancerous it is (Levin et ...engineering. Planarian worms cut into three segments “remember” which side should grow a head and a tail, and...features increased brightness and improved plasma membrane specificity. Stable Step Function Opsin - A stabilized...
  24. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...NGS assembles a larger sequence from many smaller sequences, regions with repeats may not assemble correctly...circular assembly. Most of the time, these plasmids are returned as one or two partial assemblies. If we...complete. After the run completes, we begin our assembly process. Again, thanks to our partners at seqWell...from each sample in our pool and individually assembles the reads into a single FASTA sequence that our...that are particularly difficult to sequence and assemble, including GC rich regions. For example, the CAG...
  25. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    Published
    Aug. 30, 2016, 2:30 p.m.
    ...toehold switches for Zika detection We and other members of the lab had been working on advancing the paper-based...Aedes mosquito (Messina et al, 2016). We quickly assembled a team of students, postdocs and professors to...also were careful to set up clear tasks for each member of the team to accomplish so that many elements...Sensor Constructs at Addgene As we report, we have embedded this next generation of toehold switch-based RNA...further develop our rapid prototyping and sensor assembly pipeline for future emerging pathogens or health...synthetic biology, nanomaterials science, and self-assembly for applications in health, energy, and sustainability...Listen to Our Interview with Previous Collin's Lab Member and Sample 6 Cofounder Michael Koeris Learn How...
  26. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    Published
    July 20, 2021, 1:15 p.m.
    ...permeabilize the cell membrane to allow antibodies to pass through the cell membrane. Formaldehyde fixation...degree of internal complexity such as extensive membranes produces a greater SSC.  Protein expression, measured...targets, measured by fixing and permeabilizing membranes For intracellular protein targets, users must ... dissolving lipids including those of the cell membrane. In some cases, this can mask epitopes such that...propidium iodide, can only enter cells with disrupted membranes as is the case with dead or dying cells. DNA binding...permeabilization as these methods require damaging the cell membrane and will cause all cells to stain positive. If... amine reactive dyes can only pass through the membrane of dead cells. Once in the cells, amine reactive...
  27. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...experiments: Retrieve Ensembl Transcript ID(s) of your favourite gene(s) from Ensembl genome browser: https...https://www.ensembl.org/index.html Design crRNA/ssODN: Using Ensembl Transcript ID(s) as an input query...reagents, we scaled up these tagging experiments by assembling csRNPs in a 96-well plate format and tested parallel...reagents for the knock-in experiment were pre-assembled in a 96-well plate. Glioma stem cells were transfected...be found at Desktop Genetics .For ssODN design, remember to incorporate PAM-blocking mutations (e.g. convert...in bulk and stored in small aliquots at -80 oC. Assemble csRNP: Anneal crRNA and tracrRNA using a PCR block...
  28. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...and plasma membrane reporters as well as secretory proteins or proteins targeted to membrane sub-domains...cleavage upon co-expression and spontaneous self-assembly. In an effort to obtain even more precise temporal...individual reporter constructs or as the fully assembled pMemoryArray.   Yang et al., Nat Methods 2014... Chem Biol. 2012 Apr 20;19(4):507-17. 10.1016/j.chembiol.2012.02.006. 2 Guntas et al., Proc Natl Acad...arrival in their target compartment (e.g. the plasma membrane, the extracellular space, or the lysosomes) can...plasmids with MXS-chaining. Analogous to modular assembly methods, such as BioBricks, the MXS-chaining method... organelles, a cell cycle indicator for mouse embryonic stem cells and a series of inducible promoter ...
  29. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...precursor protein. The EMBO Journal, 16(16), 5113–5122. https://doi.org/10.1093/EMBOJ/16.16.5113. PubMed ...commonly called retrovirus) family — of which HIV is a member — may seem like an unlikely candidate to use as...Figure 1 to compare the genomes of these two family members.   Figure 1: Gamma-retrovirus genome (left... RT-deficient genome. In both cases, the lipid membrane is the only component that is not dependent on...genes. It is derived from the host/producer cell membrane, and acquired as an envelope during exocytotic...E., & Teich, N. (1971). Noninfectious ARK mouse embryo cell lines in which each cell has the capacity ...
  30. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    Published
    Sept. 12, 2023, 1:15 p.m.
    ...Cloning Design primers for Gibson Assembly and NEB HiFi DNA Assembly  OligoAnalyzer Tool from IDT: For... vector database: Find sequences for plasmids assembled from publications and commercially available sources...other primer characteristics. NEB Data-optimized Assembly Design (DAD): This tool designs overhang sets ...methods, such as restriction enzyme cloning, Gibson assembly, or In-Fusion cloning.  NEB Ligation Calculator...Beth Kenkel. Additional resources were added in September 2023 by Rachel Leeson.  Additional resources...
  31. The Power Behind NGS Plasmid Validation: seqWell

    Type
    Blog Post
    Published
    April 19, 2017, 3:25 p.m.
    ...our plasmid assembly pipeline. To date, this has resulted in high quality NGS-based assemblies for over ...sequencing depth and coverage in order to, for example, assemble hundreds of different plasmids from a single Illumina...amount. plexWell™ + Addgene = high quality NGS-assembled plasmids Because of the enhanced normalization...libraries is well suited to large-scale de novo assemblies of short constructs like plasmids. The plexWell...bioinformatics pipeline that will return high-quality assembled and circularized plasmids from plexWell™ NGS data...
  32. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

    Type
    Blog Post
    Published
    June 25, 2019, 12:54 p.m.
    ... calcium chloride which permeabilizes the cell membrane so the bacteria can easily uptake your plasmid...an electrical charge to cells, to increase cell membrane permeability and thus transformation efficiency...example are bacteriophages that attach to bacterial membranes and inject their genetic material into the cell... more easily cross the negatively charged cell membrane. Physical methods such as electroporation or microinjection...microinjection actually pokes holes in the cell membrane so DNA can be introduced directly into the cell...electrical pulses to create transient pores in the cell membrane that genetic material can pass through. If you...
  33. 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.
    ...and numerous other reasons. It is important to remember that we, scientists and non-scientists alike, ...spoke in support of science - he's one of the few members of Congress who holds an engineering degree. It...the postdoctoral engagement committee and active member of the university's Postdoctoral Association.  ...career that could simply cease to exist. But then November hit and panic ensued as “alternative facts” became...encourages open discourse with our fellow community members. The March for Science was an important first step...available to people who want to join the dialog and 2) remember that science should not be partisan: everyone ...science communicator, UBC Public Scholar, and a member of the Vancouver SciCATs. Follow her on twitter...
  34. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...they cannot cross cell membranes, so they are best used either with plasma membrane proteins (such as GPCRs...fluorescence wavelengths for FRET experiments. However, remember that, if you make an FP fusion, you are inserting... Most importantly, these compounds cross cell membranes, thus enabling FRET measurements in intact cells...Intact Cells Cy3NTAc Poly His  44 35-63 Plasma membrane protein, permeabilized cells  Cy5NTAc 32 25-...Intact Cells Cy3NTAc Poly His  63 50-90 Plasma membrane protein, permeabilized cells  Cy5NTAc 43 34-...Intact Cells Cy3NTAc Poly His  65 52-94 Plasma membrane protein, permeabilized cells  Cy5NTAc 59 47-...
  35. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    Published
    Jan. 14, 2016, 3:30 p.m.
    ...freely-available DNA-editing software packages to assemble your expected full plasmid sequences (examples...computer or server. This will ensure that future members of your lab can access and use the materials they...during the cloning, sample handling, or sequence assembly steps. All of our incoming plasmids are sequenced...include cloning junctions (where most sequence assembly errors occur), deletions, mutations, in-frame ...the 5’ and 3’ ends to verify that the sequence assembly and identity of the insert are correct. In other...Primer3 to help you design your custom primers. Remember, when designing a sequencing primer, you need ...
  36. Working with Nuclear Receptors

    Type
    Blog Post
    Published
    July 15, 2014, 4:03 p.m.
    ...superfamily is comprised of 48 human and 49 murine members. NRs permit the integration and communication of...of endocrine-hormone signaling. The success of members of this group stems in part from their ability ...xenobiotic lipids derived from the diet. Additional members of this family are called orphan receptors because...coregulators exist as multiprotein complexes, a member of a single coactivator complex can serve as a ...status of SRC-3 defines its association with other members of the complex, such as p300 and CBP histone acetyltransferases...multiple cell signaling systems. Activation of membrane receptors and signaling cascades conceivably allows...
  37. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    Published
    March 23, 2021, 1:15 p.m.
    ... post: Epigenetic engineering with single-pot assembly Tracking cell-cell interactions New auxin inducible...service Epigenetic engineering with single-pot assembly By Eric Perkins Though it’s easy enough to contemplate...describes an elegant system that allows for the assembly of multiple DNA fragments efficiently and accurately... a “single pot.’   Figure 1: Single-Pot Assembly: a) Graphic depiction of chromatin within a nucleus...enzymes and reader-effector proteins. b) Golden Gate assembly (backbones available at Addgene) with examples...purposes, but the key component of the editing assemblies are sensor-actuators, either transcriptional ...
  38. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    Published
    July 23, 2015, 2:30 p.m.
    .... Since makerspaces are generally funded on a membership model - everyone contributes each month - with..., unlike your traditional biology lab, we have members with a wealth of skills to help do science on the...new replacement was not possible, but one of our members took an Arduino microcontroller and a motor controller...too expensive to make it practical - but we have members looking at how to re-use it as a platform for monitoring...fast, but with a wide range of skills among our members we’ve managed to learn a lot and cobble together... Victoria’s first hackerspace which now offers members and the public cheap access to personal manufacturing...
  39. Transferable Skills Guide: Managing a Team

    Type
    Blog Post
    Published
    Jan. 30, 2018, 2:42 p.m.
    ... There is no way around the fact that new team members need to be trained, which may seem like a waste... a seasoned report, there should be questions! Remember that delegating is not just a way to create more...for yourself, but it will help develop your team members by giving them new opportunities. I still need...comfortable with awkward silences but I’ve learned to embrace them after one ended with the person in front of...going to get you what you really want to know. Remember that one on one discussion can be confidential...Protect your team Make you team feel safe. Your team members need to trust you and you need to encourage them...
  40. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ...kinetics over 14 hours of expression in C. elegans embryos. However, this initial timer was a tetramer with...mammalian cells, in C. elegans, and in Xenopus embryos. Of course, timers may be sensitive to changes ... of vesicle fusion and release from the plasma membrane by fusing various vesicle cargoes to FP timer ...controlling pancreatic differentiation) in mouse embryos (Miyatsuka et al.) Distinguish between models for...fusing Medium-FT to LAMP-2A, a lysosome-associated membrane protein, and following its subcellular localization...with changes in gene expression. In a developing embryo, for example, a researcher could determine whether...
  41. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    Published
    March 12, 2019, 1:37 p.m.
    ...is rare these days), and a room of 5-30 faculty members. The session lasts up to an hour. You will get ...outcome with some specifics, the search committee member cannot either. Therefore, you need to be able to...idea to bounce your ideas off of senior faculty members who can help you identify overly ambitious goals...you will be interrupted frequently by committee members. As mentioned above, plan to give your 10 minute... possible to do? Research accomplishments are remembered for the problem solved, not for the amount of...faculty positions. You need to practice with faculty members that have served on search committees. If possible...
  42. Deck the Lab 2022!

    Type
    Blog Post
    Published
    Jan. 17, 2023, 2:15 p.m.
    ...pic.twitter.com/rExV3b8Q8u— Michael Früh (@Michelderapfel) December 19, 2022   Congratulations to all our #DeckTheLab...Borris Aldonza (보리스) 🇰🇷🇵🇭 (@PrecursorCell) December 12, 2022          View this post on Instagram... for either contest on Twitter or Instagram by December 31, 2022.  Follow @Addgene, use the hashtag #DeckTheLab...pic.twitter.com/rDaEjxTNZX— Heng Chooi (@YitHengChooi) December 4, 2021   I spent a year hoarding used tip boxes.../X4eKUGnQMe— Alex Dainis, PhD 🧬 (@AlexDainis) December 18, 2017...
  43. #DeckTheLab with Tinsel and Test Tubes, Fa La La La La La La La La

    Type
    Blog Post
    Published
    Dec. 2, 2021, 2:15 p.m.
    ...It’s finally December, and here at Addgene HQ in Massachusetts that means it’s time for near-freezing...decorating skills. To join the contest, simply embellish and adorn your lab space or your work from home...pic.twitter.com/OOrwo9feyJ — BTC Institute (@BTCInstitute1) December 23, 2020 Contest 2: Deck the Lab - Home Edition...pic.twitter.com/qDylF8TdHH — Matthijs Dorst (@MattNeuro) December 4, 2020 Here’s how to enter: Post your picture... for either contest on Twitter or Instagram by December 31, 2021. Let us know which contest you are entering...
  44. Five Popular Model Organisms

    Type
    Blog Post
    Published
    April 11, 2019, 12:39 p.m.
    ...an ideal model organisms for embryonic development studies as their embryos are completely transparent...., and produce 50-300 eggs at a time. Zebrafish embryos are also laid and fertilized externally, allowing... them. Scientists can simply inject one-celled embryos with DNA or RNA to edit their genomes or to create...be easily visualized in a living organism from embryonic development to adulthood. This ability to genetically... and integration of transforming sequences." The EMBO journal 10.12 (1991): 3959-3970. PubMed PMID: 1935914...
  45. Replacing Paper: Tips for Choosing an Electronic lab Notebook

    Type
    Blog Post
    Published
    July 10, 2018, 1:08 p.m.
    ...accessible to your team members only, under the specific permissions set for every member of your team. Do you...pricing, it is good to know how many of your lab members will use the ELN. If your lab is planning to grow..., notifications, sharing files, assigning team members to certain tasks, etc. How is security and user...to assign specific permissions to different lab members to avoid any issues later on. Do you need external...software solution that makes it much easier for lab members to manage and organize their work at every step...
  46. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ... Cell Free Protein Expression Kit FREQ Seq PUF Assembly Kit Below you’ll find information about what ...engages the ribosome for translational complex assembly. PhD student Dejan Gagoski used SITS to create...Academy of Sciences 109.52 (2012): 21408-21413. PUF Assembly Kit PUF proteins are composed of repeat segments...The Zhao lab has made this easy with their PUF Assembly Kit. The kit contains all the variant PUF segments...Sequence‐Specific Detection of RNAs in Two Colors."ChemBioChem 17.10 (2016): 895-899. Choudhury, Rajarshi,...15875. See Kendall Morgan’s blog post on the PUF Assembly Kit for some insights on its potential from its...
  47. Designer PUF Proteins for Any RNA Target

    Type
    Blog Post
    Published
    June 17, 2014, 7:57 p.m.
    .... Get the PUF Assembly Kit! Tips and tricks For those interested to try the PUF Assembly Kit out for themselves...tool…designer PUF proteins! A newly available PUF Assembly Kit makes it possible to devise RNA binding proteins...factors (PUF). In a single step, researchers can now assemble designer PUF domains for RNA specificity engineering... Reference Abil Z, Denard CA, Zhao H.  Modular assembly of designer PUF proteins for specific post-transcriptional...
  48. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    Published
    Oct. 17, 2019, 12:28 p.m.
    ...flow of electric charges across cell membranes and maintain membrane potential in response to visible light...across the axon's membrane: once a certain number of positive ions crosses the cell membrane, a threshold ...rhodopsins enable ions to flow across the axon’s membrane, thereby controlling neural activity (Bernstein...Stoeckenius. "Rhodopsin-like protein from the purple membrane of Halobacterium halobium." Nature new biology...
  49. Viral Vectors 101: Parts of the AAV Packaging Plasmid

    Type
    Blog Post
    Published
    Dec. 3, 2020, 2:15 p.m.
    ...also encodes the Assembly-Activating Protein (AAP) and the recently discovered Membrane Associated Accessory... AAV genome and docks with the genome onto an assembled capsid. Then Rep52/40, which also bind the capsid...generate the three VP proteins. The three VP proteins assemble into capsid at a 1:1:10 ratio (VP1:VP2:VP3). There...reading frame (ORF). While AAP promotes virion assembly, the role of MAAP has not been clearly defined...
  50. FlipGFP, a novel fluorescence protease reporter to study apoptosis

    Type
    Blog Post
    Published
    May 21, 2019, 12:10 p.m.
    ...apoptosis reporter, ZipGFP, which utilized the self-assembling split GFP (GFP1-10 and GFP11) to increase fluorescence...animals. To do this they modified a different self-assembling split GFP (GFP1-9 and GFP10-11) by flipping a...rectify this the Shu lab first improved the self-assembling capability of sfCherry approximately 20 fold ...FlipGFP to study apoptosis in HeLa cells, zebrafish embryos, and Drosophila. They treated HeLa cells expressing...the spatiotemporal dynamics of apoptosis during embryo development. GFP fluorescence increased over time...
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