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We narrowed to 810 results for: nin

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  1. Running for Rare Disease, Running for FOP, Running for AJ

    Type
    Blog Post
    Published
    Sept. 15, 2016, 2:30 p.m.
    ... my poor, poor legs. Running for FOP, running for AJ Runners for #Running4Rare are paired with patient... I learned there was a Genzyme-based running team called Running for Rare Disease that runs on behalf ...mall. I took action, joined a gym, and started running on a treadmill. After being with the company for...parents, Kristi and Rico, had to keep him from running too much as any jarring of his body could initiate...raising funds and awareness of FOP. Please consider running with us or making a donation via the link below...the emotion HOPE; a fitting superpower for those running on behalf of rare disease sufferers like AJ. As...development for First-In-Man studies. Kurt has been running on behalf of AJ Gonzales, a young boy suffering...

  2. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    Published
    Nov. 1, 2016, 2:30 p.m.
    ...these include, TA cloning, ligation independent cloning, TOPO cloning, one step cloning, and overlap extension...Standard Restriction Cloning and Fast Cloning Requirement Restriction Cloning Fast Cloning Choose Restriction...restriction cloning is still routinely performed by many labs, a variety of other cloning techniques with...excessive time requirements. FastCloning is a simpler yet reliable cloning technique that was developed... Transformation + + The FastCloning process in brief In FastCloning, you PCR both the vector and ...and ligated in vivo. Detailed FastCloning procedure To start the FastCloning process, the insert and vector...the multi-cloning site, and the reverse primer is located in the 5’ end of the multi-cloning site. The...

  3. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    Published
    Sept. 15, 2020, 1:15 p.m.
    ...a look at a few screening methods. Blue-white screen A widely used method of screening is the blue-white...biology, cloning a gene into your plasmid is often a numbers game. By using a variety of screening and selection...If you’re cloning a plasmid, you’ll need a way to find the needle in the haystack: the one perfect clone...use positive and negative selection in plasmid cloning.  But oftentimes, a selection strategy is not ...desired plasmid. In many cases, you will need a screening strategy as well. In a screen, you aren’t killing...selection, then screen Why add a screen to your cloning strategy? A screen will help you more easily identify...resistance marker in the backbone. A selection on agar containing ampicillin will yield bacteria that have taken...

  4. MXS Chaining

    Type
    Blog Post
    Published
    Feb. 7, 2017, 3:30 p.m.
    ...directional cloning   The principle behind MXS-chaining is repeatable, restriction-enzyme based (chaining-based...between various cloning methods are listed in Table 2. For example,  chaining-based cloning methods require...various chaining-based cloning methods are listed in Table 3. The goal of high throughput cloning is to ....org. Table 3: Chaining Based Cloning Methods Method Restriction Enzymes Chaining Direction MXS-Chaining...a useful application of high-throughput cloning. MXS Chaining is one such technique and has been used ...Neveu, 2015). Origin and purpose of MXS chaining MXS-chaining was designed to create plasmids for fluorescence... generated MCS. MXS-chaining also has the advantage of being directional, meaning that (in the previous...

  5. Plasmid Cloning by PCR

    Type
    Blog Post
    Published
    March 29, 2016, 2:30 p.m.
    ...plasmid. Designing primers for PCR based cloning     The basic PCR primers for molecular cloning consist...outlined in our restriction cloning post with a few quirks specific to the PCR cloning process - please check...our restriction cloning post. However, even more than with standard restriction cloning, it is essential...In its simplest form, PCR based cloning is about making a copy of a piece of DNA and at the same time...Restriction Site: Your chosen restriction site for cloning (usually 6-8bp) Hybridization Sequence: The region...your recipient plasmid (usually in the Multiple Cloning Site (MCS)), but do not cut elsewhere on the plasmid...these are used for primer design: Because we are cloning an ORF, we want to clone from the start codon (...

  6. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    Published
    Jan. 12, 2017, 3:30 p.m.
    ...attL-entry-TOPO vector. TOPO cloning adds short end(s) to facilitate cloning into an attL-containing entry vector.   ...TOPO cloning the insert into an attL-containing entry vector.   Method C: Restriction cloning of a...facing a cloning project, scientists are no longer limited to traditional restriction enzyme cloning. Instead...Gateway cloning method and its advantages. An introduction to Gateway technology The Gateway cloning method...PCR product with an attP-containing donor vector.   Method B: TOPO-cloning of the desired insert into... is inserted in a multiple cloning site (MCS) of an attL-containing entry vector.   Figure 5: ...create an entry clone: Restriction cloning the inset into an attL-containing entry vector.   Pro Tip: ...

  7. Plasmids 101: Modular Cloning

    Type
    Blog Post
    Published
    March 12, 2024, 1:15 p.m.
    ...strangers to cloning. If you’re reading this, then chances are you have been in the cloning trenches yourself...yourself. Modular cloning, or MoClo, is a systematic cloning strategy designed by synthetic biologists...experiments it enables. What is modular cloning? Modular cloning, or MoClo, is an organization scheme for... MoClo reactions use Golden Gate cloning, not all Golden Gate cloning is MoClo. That’s because MoClo specifically...biologists to perform large scale, one-tube cloning reactions. In this blog, we’ll cover the basics of what MoClo...hierarchy. MoClo originally developed from Golden Gate Cloning, in which a type IIS restriction enzyme and a ligase...schema described above, and not any particular cloning technique. Though rarer, you can find MoClo that...

  8. Plasmids 101: TOPO Cloning

    Type
    Blog Post
    Published
    Oct. 27, 2016, 2:30 p.m.
    ...Topoisomerase based cloning (TOPO cloning) is a DNA cloning method that does not use restriction enzymes...TOPO-TA) cloning; however, the TOPO cloning technique has also be adapted for blunt end cloning. So how...researcher to perform blue-white screening after transformation - self joining of the vector ends results in... of TOPO cloning happens. Basic procedure Let’s break down the steps needed for TOPO cloning: 1. Create...on the basic ability of complementary basepairs adenine (A) and thymine (T) to hybridize and form hydrogen...how does TOPO cloning work? As illustrated in the Figure below, the “A” overhang on the blue PCR product... endonuclease and as a ligase by cleaving and rejoining supercoiled DNA ends to facilitate replication...

  9. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    Published
    Oct. 18, 2018, 12:37 p.m.
    ... bacterial in vivo cloning is now able to come of age. The in vivo assembly cloning protocol In Vivo Assembly... to drive assembly. Combining multiple modifications A key feature of IVA cloning is the ability to combine...assembled using IVA cloning, although the efficiency of assembly decreases as the cloning complexity increases...Watson, and Ingo H. Greger. "IVA cloning: a single-tube universal cloning system exploiting bacterial in...before choosing a cloning method Check out all of our blog posts on plasmid cloning Resources on Addgene.org...the MRC Laboratory of Molecular Biology. Plasmid cloning is an essential part of any molecular biology project... experimental process. The majority of current cloning techniques involve the assembly of a circular plasmid...

  10. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    Published
    Feb. 18, 2016, 3:42 p.m.
    ...Transformation containing the ligation mixture with backbone alone; 2. Transformation containing ligation mixture...When cloning by restriction digest and ligation, you use restriction enzymes to cut open a plasmid (backbone... digests.   Overview of the restriction cloning process. Both the plasmid (blue, backbone) and ...the insert and backbone covalent.  Before beginning the restriction digest and ligation process, you... proper orientation. For instance, if you were cloning a gene into an expression vector, you would want...downstream of the promoter as part of a multiple cloning site (MCS). Having multiple sites allows you to...it via their different migration speeds; after running the gel for some duration of time, these differently...

  11. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    Published
    Sept. 24, 2019, 12:55 p.m.
    ...the steps in the cloning process, there are also many ways to troubleshoot the cloning experiment. Here...a different cloning strategy. Once you've identified potential clones after the screening, sequence the... DNA Custom Cloning Service since 2009. He has  developed a proprietary, low cost cloning protocol that... contributed by Oskar Laur, head of the custom cloning core at Emory University, and Paolo Colombi, a ...a product development scientist at Addgene. Cloning can be quite an arduous process. The PCR could fail...Here are some tips that will help you with your cloning project, and hopefully obtain your coveted plasmid...diving into the experimental work, spend some time outlining the construction of the plasmid and all the steps...

  12. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    Published
    Sept. 7, 2017, 12:17 p.m.
    ...dried out and died. The alternative to cloning rings, dilution cloning into 96-well plates, is extremely time... to the expense of dilution cloning. Taking a page from bacterial cloning, we reasoned that we should...Tips for using the agarose cloning method Test your agarose temperature. Cloning cells using an agarose overlay... laboratory became frustrated with using glass cloning rings to isolate colonies of antibiotic-resistant...time required to grease, place and fill a dozen cloning rings, the remainder of the colonies on the plate...thin layer of molecular biology-grade agarose containing physiological concentrations of salts and glucose...glucose. If we circled well-separated clones containing more than 50 cells (visible to the naked eye) with...

  13. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    Published
    Aug. 27, 2015, 2:30 p.m.
    ...Advantages of Golden Gate cloning Golden Gate cloning is one of the easiest cloning methods in terms of hands-on...enzyme-based cloning methods. Each method has its own pluses and minuses, but Golden Gate cloning has been... and easy-to-use system to your cloning efforts. Golden Gate cloning technology relies on Type IIS restriction...construct, allowing for precise, scarless cloning. The cloning scheme is as follows: the gene of interest... leveraged the power of Golden Gate cloning into a modular cloning strategy. Sometimes referred to as ...target sequence into a Cas9-containing plasmid such as pX330. This cloning strategy not only makes it ...easy.  Disadvantages of Golden Gate cloning Golden Gate cloning is not 100% sequence-independent: to ...

  14. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ...and ease of generating gRNA-containing plasmids. CRISPR genome wide-screening experiments commonly use lentivirus...means to carry out so-called “forward genetic screening experiments”, it is certainly the most robust....repaired through error-prone non-homologous end joining (NHEJ), usually results in a loss-of-function mutation...libraries are composed of many different gRNA-containing plasmids, each with a different genomic target...genome-wide screens? The goal of a genome-wide screening experiment is to generate and screen a population...CRISPR can be readily scaled up for genome-wide screening due to the broad range of potential target sequences...consist of a heterogeneous population of gRNA-containing lentiviral transfer vectors, each targeting a...

  15. Plasmids 101: Blue-white Screening

    Type
    Blog Post
    Published
    June 4, 2015, 1:03 p.m.
    ...insert (blue-white screening). Other types of screening methods Although blue-white screening is probably the...talk about an even more specific method of screening your cloning reaction. Being able to select for colonies... that! (To read about other screening strategies, check out our screening overview blog post.) *Note:...Blue White Screening Available at Addgene Are: pUC18 and pUC19 Let’s begin at the beginning. The well-...plasmid, creating an α-complementation cloning vector. When a cloning reaction goes to plan and your DNA ...Blue-white screening only indicates the presence of AN insert, not necessarily YOUR insert. Any cloning artifact... few common vectors. Blue-white screening is just that – a screening process. It does not select only...

  16. Finding and Joining Your Dream Lab

    Type
    Blog Post
    Published
    April 8, 2014, 2:50 p.m.
    ... many more suggestions on planning for future success by finding and joining your dream lab, watch the... joining your dream lab...

  18. Learning Technical Skills at a Lab Central Workshop

    Type
    Blog Post
    Published
    Feb. 20, 2018, 2:30 p.m.
    ...I understand? How does the information I’ll be learning relate to me as an individual and Addgene as a...constantly update our processes and get new ideas from learning experiences like workshops at Lab Central. Because...

  20. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    Published
    Nov. 14, 2017, 2 p.m.
    ...that many researchers feel that they lack formal training in a variety of transferable skills. At Addgene... where you might need some help. Today: Career Planning Resources. The key to ensuring success during ...any formal or mandatory professional development training. Further, even for those students and researchers... page, where you’re likely to find current job openings. A good approach is to do this first for companies...way, once you’ve identified an appropriate job opening, you can send your contact an email with the relevant... graduate student or post-doc in a traditional training program, and you hope to make a career in academia...own Joanne Kamens, a strong advocate for career planning, has written a great blog on choosing good mentors...

  21. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    Published
    Feb. 13, 2025, 2:15 p.m.
    ...deaminases. Adenine base editors The beginnings Nicole Gaudelli from David Liu’s lab created an adenine base ...cytosines preceded by guanine (Thuronyi et al., 2019). Following the development of adenine base editors (discussed...Creating an adenine base editor required an additional step because there are no known DNA adenine deaminases...about combining the function of both editors into one? This has been done by fusing the adenine and cytosine...cytosine and adenine base editors Grünewald et al., 2020 SPACE   Fused cytosine and adenine base editors...by researchers. Compared to non-homologous end joining (NHEJ), HDR occurs at a relatively low frequency... editors were cytosine base editors (CBEs) and adenine base editors (ABEs). CBEs mediate a C to T change...

  22. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    Published
    May 14, 2024, 1:15 p.m.
    ...another great option for mammalian cloning, and is geared towards cloning multi-TU expression constructs ...blog Plasmids 101: Golden Gate Cloning High Complexity Golden Gate Cloning with NEB’s DAD New MoClo basics...Modular cloning is a popular DNA assembly tool used to build single- and multi-gene constructs for a ...goals are more general in nature, or you aren’t planning on using K. phaffii, then the MoClo YTK Kit specifically...should also take a look at the Yeast GoldenBraid Cloning System (Pérez-González et al., 2017). In addition... kit is a great starter for your general plant cloning experiments. Addgene also has two additional MoClo... kit is also compatible with the Joint Modular Cloning Kit which it builds upon (its ‘basics’ MoClo plant...

  23. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    Published
    March 30, 2017, 2:30 p.m.
    ...in a sample. Digital PCR assays for mutation screening Now that we have introduced how digital PCR works...indel mutations resulting from non-homologous end joining (NHEJ) (2-5), depending on your desired genome ...the bi-allelic mutations typically desired when screening clones for complete functional knockout of your...interested in harnessing the power of digital PCR screening to validate your genome edits, this paper provides...Remember that although digital PCR is a great screening tool, it still requires target DNA sequencing ...PCR-Based Method for Efficient and Highly Specific Screening of Genome Edited Cells. PLoS ONE 11, e0153901–...

  24. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    Published
    April 12, 2016, 2:30 p.m.
    ...multiplexed CRISPR systems for high-throughput screening are vastly useful for mapping the combinatorial...ligation steps to build barcoded lentiviral plasmids containing one or more gRNAs. As each ligation reaction ...digestion and ligation result in plasmid libraries containing combinations of different gRNAs in each plasmid... lentiviral vector backbone pAWp12 (Figure 2). Cloning from the pAWp28 storage vector to the pAWp12 lentiviral...library, but this should be verified by NGS before beginning your experiments. Figure 2: Strategy for...Hiroko, et al. "Genome-wide recessive genetic screening in mammalian cells with a lentiviral CRISPR-guide...Ophir, et al. "Genome-scale CRISPR-Cas9 knockout screening in human cells." Science 343.6166 (2014): 84-87...

  25. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...your rate of interactions when screening! Tips for two-hybrid screening Minimizing false positives: Run... these discoveries to create a new system for screening protein-protein interactions. In later years, ...reporter gene cloned downstream of a promoter containing the target sequence of the DBD. The DBD-POI fusion...protein interact and enable intein splicing (5). Screening with yeast two hybrid systems While initial studies...plasmid transformants into a diploid strain for screening), or arrayed on a 96 well plate where each well...a defined prey fusion.  The former allows for screening greater numbers, but the latter eliminates the...the step of identifying prey proteins after screening. It is also possible to screen libraries of bait ...

  26. Gaining Leadership Skills Volunteering at a Professional Organization

    Type
    Blog Post
    Published
    Aug. 6, 2015, 2:30 p.m.
    ...should acquire during their PhD and postdoctoral training. Addgene executive director Joanne Kamens has ...scientific society in your field. My leadership training began with the NPA where I gained invaluable experience...function without long distance communication and learning to work with people despite the distance improved...leadership institute’ weekends would entail intense training to develop our leadership and management skills...junior leaders what I had experienced and learnt. Opening doors to further opportunities  Having gained leadership... students to think about their career options, gaining transferable skills such as leadership skills can...

  27. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    Published
    April 16, 2015, 3:45 p.m.
    ...two major repair pathways – Non-Homologous End Joining (NHEJ) and Homology Directed Repair (HDR) – and... pathways work, as this could be relevant when planning your experiment. Here, we’ll discuss NHEJ, and...mediated DSBs in the genome. Non-homologous end joining steps Unlike HDR, NHEJ is active throughout the...nucleases (Artemis), and structure-specific end cleaning enzymes (Aprataxin, Tdp2) that function to make... breaks, and could result in deletion of the intervening DNA (“pop-out” deletions) if NHEJ joins the distal...repair (“pop-in” insertion) provided a template containing compatible overhangs is available. Cas9 can also...David Wyatt is a graduate student interested in determining how the structure of broken ends impacts how ...

  28. Future of Research Conference - Remarkable Opening Session

    Type
    Blog Post
    Published
    Oct. 7, 2014, 3 p.m.
    ...such as new training programs (like at the Keck Graduate Institute), funding for training (such as the...speaking out and writing about issues in our science training infrastructure for some years.  He spoke eloquently... to choose institutions with allies engaged in training excellence to begin effecting change.  Panelists...advisors only know about academia and alternate training needs to be provided to take advantage of all ...the conference organizing team.  The topics were Training, Structure of the Workforce, Metrics and Incentives...Scientists must do science to be happy so the beginnings of solutions can’t come too soon.  Watch the ...

  29. Developing Transferable Skills During Science Training

    Type
    Blog Post
    Published
    June 3, 2014, 2:14 p.m.
    ...transferable” skills you have developed in addition to learning how to be a scientist. Why not stack the deck ...skills in ways that will also enhance your science training and that will put you in position to pursue a ...develop those transferable skills while you are also learning to be an excellent scientist. Leadership and management...association) Collaboration and teamwork One of the defining characteristics of nonacademic science careers... science training...

  30. A Guide to Designing a Scientific Poster: Content, Layout, and More

    Type
    Blog Post
    Published
    Oct. 22, 2019, 12:20 p.m.
    ... to help you build a great poster.  Tips for designing your poster Check conference guidelines To begin...you should always follow conference rules when designing your poster, since they can differ. 48” x 36” ...

  31. Plasmids 101: Positive and Negative Selection for Plasmid Cloning

    Type
    Blog Post
    Published
    Aug. 22, 2019, 12:43 p.m.
    ...themselves. Therefore, your cloning strategy needs to identify cells containing the plasmid construct you...positive selection, negative selection, and/or screening. We’re focusing on positive and negative selection...After transformation, plate the cells on medium containing the corresponding antibiotic. For example, if...cassette, you would plate the cells on kanamycin-containing agar. Only cells carrying plasmids that contain...with or without your inserted DNA) and further screening will help you determine if the plasmid backbone...Auxotrophy and prototrophy come into play during cloning if you’ve designed your experiment using an auxotrophic...amino acid). Many common E. coli strains used for cloning are auxotrophic for specific amino acids. When ...

  32. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    Published
    March 12, 2019, 1:37 p.m.
    ...Hughes Medical Institute. Are you currently on or planning to go on the academic job market? In addition ...-10 minutes to present your research vision uninterrupted and then you will begin working through your...overview and address the aims in more depth in the remaining time. The typical chalk talk structure is: What...

  33. From Lab Bench to Leadership: Transitioning into Management as a PhD

    Type
    Blog Post
    Published
    Jan. 21, 2025, 2:15 p.m.
    ... most pivotal decisions many PhDs face when transitioning from academia to industry is whether to stay...equips you with exceptional technical skills, transitioning into management often means stepping into unfamiliar...less time than the seemingly burdensome task of explaining the steps to a lab assistant. However, I recognized... member has unique motivations, strengths, and learning styles. For example, during my time as a lab scientist...and skills that were quite different from mine. Learning to bridge those differences was a pivotal moment...Finally, developing emotional intelligence is vital. Learning to "read the room," understanding team motivations... programs on platforms like Coursera, LinkedIn Learning, or edX. Podcasts: HBR IdeaCast Coaching for...

  34. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ...If cloning methods had personalities, SLIC (sequence- and ligation-independent cloning) would be a true...starting point: Ligation-independent cloning Ligation-independent cloning (LIC) was first developed in the... doing large amounts of cloning. Assembly is scarless, unlike Gateway cloning, and the method’s flexibility...Ligation-Independent Cloning Protocol Gibson Assembly Blog Post Choosing a Molecular Cloning Technique Choosing...the 1990s. While traditional restriction enzyme cloning uses short sticky ends to insert genes into plasmid...bacteria after transformation. LIC is a reliable cloning method, but it is limited by its sequence constraints...designed plasmids. Sequence- and ligation-independent cloning: A SLIC makeover In 2007, LIC received an important...

  35. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    Published
    May 28, 2024, 1:45 p.m.
    ..., we'll discuss the principles and process of designing a flow panel.  Principles of excitation and emission...1 shows a graph from the BD® Spectrum Viewer, outlining a few commonly used fluorophores and their emission... them. Alright, let's build your first panel! Designing a Panel In this example, I will describe a basic...Besides some background cell death constantly happening in cell culture, the transduction process itself...through each section of this article again. In the beginning, we talked about the relationship between a photon's...

  36. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ...deep mutational scanning.     3 reasons to be interested in deep mutational scanning Hundreds of thousands...Scientists use deep mutational scanning to simultaneously test how multiple amino acid changes affect... to phenotype that can be used with mutational scanning: ligand binding assays, phage display assays, ...Several methods can be used to create mutational scanning libraries. While each of these methods works well...fragments in a single-tube reaction. For mutational scanning experiments, libraries of DNA fragments that contain...assembly is better suited for creating small, focused scanning libraries. PFunkel: Of prior existing comprehensive...Variants on a Large Scale Using Deep Mutational Scanning.” Nature Protocols 9.9 (2014): 2267–2284. PubMed...

  37. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    Published
    Dec. 8, 2016, 3:30 p.m.
    ...Plasmids 101: TOPO Cloning Plasmids 101: Golden Gate Cloning Read all of our Plasmid Cloning Posts Resources...more recently developed Gibson Assembly Cloning and Gateway® cloning technologies, as well as several others...powerful cloning tool Molecular biologists first saw the potential of this system for enhancing cloning efficiency... prep plasmids containing the ccdB gene, which can then be used for downstream cloning applications. While...strains for more strain info. Additional cloning resources As cloning continues to evolve, understanding all...Addgene.org Choose a Molecular Cloning Technique Find Plasmid Cloning Protocols Check Out All of Our ...If you’re into cloning, you’re probably aware that there are several methodologies currently available...

  38. Plasmid Grant to Aid Cervical Cancer Screening in Ghana

    Type
    Blog Post
    Published
    Nov. 8, 2016, 3:30 p.m.
    ...Cervical cancer screening in Ghana Gladys is currently working to enhance cervical cancer screening in Ghana....cervical cancer screening for a limited group of Ghanaian women by including PCR screening for HR HPV alongside...tests are commonly used in Ghana, there is little screening for DNA markers like the presence of high risk...tests currently in use. Gladys will use plasmids containing a variety of HPV genes as positive controls in...by these screens can then undergo more routine screening allowing for early cervical cancer detection, ...her in the future. Future awardees We will be opening the Addgene/Seeding Labs Plasmid Grant to a third... cancer screening...

  39. IDP and your PI: A Roadmap for Career Planning and Personal Development

    Type
    Blog Post
    Published
    July 21, 2015, 2:30 p.m.
    ...development through thoughtful planning and honest feedback DePace’s yearly planning meetings revolve around ...various issues, the PI is responsible for determining meaningful solutions. PI-trainee communication builds...setting and career planning help alleviate the anxiety of graduate and postdoc training. On the other side...goals.      Although myIDP is great for career planning, it shouldn’t be used in isolation. Without clear... her lab (pictured above). DePace holds yearly planning meetings with each trainee to create a supportive...to facilitate honest conversation. Goals and Planning Worksheet: Lists accomplishments, research/professional...Starting with a recap of accomplishments, from learning new techniques to finally figuring out a tough...

  40. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

    Type
    Blog Post
    Published
    March 17, 2020, 1:32 p.m.
    ...and base changes from A•T-to-G•C are mediated by adenine base editors (ABEs). How does this work? Through...dilution (passage from the infected to a fresh uninfected host cell culture). This dilution step represents...passaged from an infected host-cell culture to an uninfected culture which results in less overall dilution...BioRender.com. Introducing ABE8e - a fast and flexible adenine base editor Lower-stringency PANCE, followed by...efficient in on-target DNA editing as ABE8e while maintaining the lower off-target DNA and off-target RNA editing...pathogenic point mutations. Together, cytosine and adenine base editors can correct all four of these kinds..., Liu DR (2020) Phage-assisted evolution of an adenine base editor with improved Cas domain compatibility...

  41. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    Published
    Sept. 26, 2019, 12:50 p.m.
    ...they think of these particles as tiny DNA cutting ninjas that deliver CRISPR to multiple types of targets...fluorophore or tag. To utilize HDR, a donor template containing the desired sequence flank by homology arms must...hydroxyphenylpyruvate dioxygenase (HPD) gene, all nine treated mice had ~10% editing in their livers, while...line on Nanoblades Nanoblades are tiny DNA-cutting ninjas which can stealthily delivery many types of CRISPR...

  42. Making Career Development an Integral Part of the Science Training System

    Type
    Blog Post
    Published
    Feb. 25, 2021, 2:15 p.m.
    ...that career skills training is a required and integral part of all science training programs. I believe...the transition to a post-academic career. This training was “included” with their tuition, so to speak...even doing a very good job of providing skills training for scientists destined to continue in academia... science trainees to be successful after their training.   Figure 1: Professional Development Hub...driving systemic change in academic career readiness training. It's been an honor to work with such a group ...know-how to integrate career development in science training programs. In 2018, the ASBMB and Burroughs Wellcome...representatives from all areas of the scientific training and employment infrastructure. A Stakeholder Workshop...

  43. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

    Type
    Blog Post
    Published
    Feb. 20, 2014, 4:57 p.m.
    ...to enable both negative and positive selection screening. They and their colleagues including Feng Zhang... (PubMed). Genome-Scale CRISPR-Cas9 Knockout Screening in Human Cells. Shalem O, Sanjana NE, Hartenian...Dec 12. (PubMed). Genome-Wide Recessive Genetic Screening in Mammalian Cells with a Lentiviral CRISPR-Guide...

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

    Type
    Blog Post
    Published
    Aug. 21, 2018, 12:31 p.m.
    ...limited - what cloning approach should you use if the budget is particularly tight? Most cloning processes ...for traditional restriction cloning. Sequence and Ligation Independent Cloning (SLIC) could also be a great... The general Gateway cloning protocol is fairly flexible and adaptable to cloning different types of DNA...guarantees, some cloning techniques do tip the scales in your favor. For starters, Golden Gate cloning is extremely...since CRISPR; you are ready to clone! But...how? Cloning is no longer limited to restriction enzymes - although...: Speed Cost Difficulty Efficiency For cloning in particular, another important consideration ...inserts ...as some methods are more amenable to combining multiple DNA fragments than others. In today’s...

  45. BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development

    Type
    Blog Post
    Published
    June 26, 2018, 1:33 p.m.
    ...advance their research on diseases of glycosylation, arginine mutagenesis, and neural development. Paula Videira...inherited metabolic diseases with severe and life-threatening consequences. They result from defects in different...drugs can be used to treat these diseases (drug repositioning). The ultimate goal of this work is to lower...PubMed PMID: 27393411. Michael McMurray: Rescuing arginine mutations in rare disease with guanidine Michael... mutations that change protein sequence affect arginine (Arg) amino acids more frequently than any other...

  47. Addgene’s Newest Way to Speed Your Research: The Cloning Grade DNA Service

    Type
    Blog Post
    Published
    July 28, 2020, 1:15 p.m.
    ...some of them containing fluorescent markers or protein tags, and suitable for molecular cloning. Plus, we...bacteria and prep the plasmid before you begin your cloning experiment? What if after receiving the plasmid...collection has been prepared as part of our new cloning grade DNA service and is ready to use out of the...the box. We estimate that providing cloning grade DNA could save scientists up to three days of work. ...discovery.” Here are the details. Plasmids offered as cloning grade DNA As we begin testing out this service,...directly used in the research on the SARS-CoV-2. Nine of the overall plasmid selection will be open to...of target organisms (from bacteria to mammals), cloning techniques, and experimental use (CRISPR, viral...

  48. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ...Adenine base editors (ABE) mediate A•T-to-G•C base changes (Figure 1), but it can be challenging to make...et al., 2020. Directed evolution identifies adenine base editors with improved editing in cell lines...TadA sequences to create the eighth generation of adenine base editors, called ABE8 (Gaudelli et al., 2020...Cas9 proteins to increase the targeting scope of adenine base editors The most commonly used S. pyogenes...(“N” can be any nucleotide base, followed by 2 guanines). This limits the use of ABEs as DNA sites that...off-target RNA and gRNA-dependent DNA editing while maintaining on-target base editing functionality.  The delivery... LE, Ciaramella G (2020) Directed evolution of adenine base editors with increased activity and therapeutic...

  49. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

    Type
    Blog Post
    Published
    Feb. 18, 2020, 2:15 p.m.
    ...Iterative improvement - Making Binning Singletons better Our experience running this program at ASM Microbe...This post was contributed by Joe James from Binning Singletons. The sheer scale of a large conference...bench scientist working at an institution where training opportunities weren’t always available. Even more...Denise Akob and Geof Hannigan that the outline for Binning Singletons came together. The term Singleton was...any reason was a Singleton. Figure 1: The Binning Singletons logo. Image: James et al., 2020. ...similar sequences together in bins, we thought that binning Singletons together would provide a structure and...Singletons in their bins.  The next component of Binning Singletons is when we bring all the bins together...

  50. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    Published
    March 1, 2016, 3:30 p.m.
    ...circular polymerase extension cloning) and SLiCE (Seamless Ligation Cloning Extract) to name two more. Likewise... Assembly (NEB) Cold Fusion Cloning (System Biosciences) In-fusion Cloning (Clontech) Although these ...smaller fragments. Know your cloning methods There are two other common cloning methods that are easy to ...different methods. Golden Gate cloning does result in the seamless joining of fragments, but uses site specific...your insert. Gateway cloning utilizes λ integrase to catalyze directional cloning of DNA parts that are..., Gateway, or Golden Gate cloning methods. Whether it is for routine cloning, assembly of multiple fragments...About Other Cloning Techniques Additional Resources on Addgene.org Find Plasmid Cloning Protocols Find...
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