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Showing: 281 - 300 of 756 results

  1. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    Published
    Nov. 1, 2016, 2:30 p.m.
    ...purification before transformation. After transforming cells, you should perform colony PCR with vector..., ligation, and transformation. While restriction cloning is still routinely performed by many labs, a...products are then transformed directly into chemically competent E. coli and transformants selected with ...+ + Gel Purification + - Ligation + - Transformation + + The FastCloning process in brief In ...for this procedure to work properly. Directly transform chemically competent cells with the digested mixture...-cycle PCR leads to the most colonies after transformation. When a 1:1 vector:insert ratio is used in ... and 2 µL of digest product are used in the transformation, >70% of the colonies are correct. Good luck...

  2. Management for Scientists: Giving Feedback

    Type
    Blog Post
    Published
    Feb. 6, 2015, 2:46 p.m.
    ...AMA   Guidelines for giving feedback Use formal and informal mechanisms for feedback (more on this below...her management style. Formalizing the feedback session Giving regular, informal feedback is a very effective...Using this standard format not only enables you to discuss a person's performance or what has been learned...scientists seem to find it easier to give feedback in formal sessions using a specific list of questions after... it helps to plan effective next steps. For a formalized feedback session, answer the questions below ...discuss openly. Both parties get used to using this format and it becomes even more effective over time if...of indirect language You must be honest about performance problems. While giving corrective feedback isn...

  3. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    Published
    Nov. 3, 2016, 2:30 p.m.
    ...organizational performance and often have to be able to quickly synthesize information from a variety ... their visual format is quite appealing to many audiences. While all of these platforms were developing...not hosted on the platform, they can be hosted on YouTube, Vimeo, and other platforms (such as university...quickly. Taking what I loved from each of these platforms (such as social networking, visual appeal, instant... across multiple other sites), I created a new platform for sharing video abstracts about research: www.WeShareScience.org...www.WeShareScience.org. WeShareScience is a free platform for sharing short, 3 to 5 minute videos about ...university websites). WeShareScience is a social platform that provides links to the videos, keeping the copyright...

  4. Behind the scenes: Addgene’s new search engine and more

    Type
    Blog Post
    Published
    Dec. 12, 2019, 2:22 p.m.
    ...The ability of our users to discover all this information, however, did not improve at the same pace as...as the quantity and quality of the information. As a result, resources on the website were hard to find... than that: the general “findability” of the information on the site. Different users have different strategies...paths, the organization and completeness of our information, and the accuracy and relevance of our search...the advantage of a very specialized domain of information so we were able to fine tune our approach much...than if we were dealing with general purpose information. Below are a few of the strategies we used in...Categories We created a categorization of our information in order to more granularly customize the search...

  5. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    Published
    Nov. 16, 2021, 2:15 p.m.
    ...optimization as each antibody performs differently and some antibodies impact the performance of other antibodies...beautiful results. Performing an immunofluorescence experiment The process to perform IF can be broadly...important considerations to make when planning and performing an IF experiment. What is immunofluorescence... different species and colors makes it easy to perform multiplexed IF. By using multiple primary antibodies...in IF experiments. Precisely which controls you perform will depend on your experimental context.  Choosing... to use, then the primary antibody will likely perform as expected in both.  How do you find validated...may be, it is always important to verify their performance in your hands and in your specific experiment...

  6. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    Published
    Feb. 7, 2019, 2:16 p.m.
    ...80) are transformed to 100. This transformation (illustrated in the graph below) is performed because ...order reagents Perform your wet lab experiment Analyze the data For more detailed information on using CrispyCrunch...even more.) However, none of the tools we found performed batch guide design, batch primer design or batch... burden of having to manage all the standard information that goes into a 96-well plate and comes out ... reports. With CrispyCrunch, you get the all information from Crispresso plus a summary per batch. Ranking... at a genome by eye. We came up with a ranking formula that does just this: Rank = (compressed CFD score...the DNA target. CFD scores are then modified (“transformed”) such that weak interactions (e.g., scores <...

  7. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    Published
    Jan. 3, 2019, 1:35 p.m.
    ... base editing platforms for plant systems.  Figure 2: Plant base editor platforms and their molecular...72 hours post-transformation/transfection in a transient assay. For stable transformation, regenerated ... reducing the likelihood of detrimental indel formation. After binding with DNA, the CBE deaminates a ...G)? You may wonder if a CRISPR-base editor can perform this change. However, there are no naturally known...deaminases (ecTadA), fused it with dCas9, and transformed it into E. coli harboring the defective CamR ...genome edited plants via Agrobacterium-mediated transformation. We also constructed smaller vectors suitable...suitable for transient assays using protoplast transformation. Cytosine base editors for plant There are several...

  8. Summer SciComm Series: Cognitive Load

    Type
    Blog Post
    Published
    June 16, 2022, 1:15 p.m.
    ...to read a formulaic fiction story that doesn’t require any detailed retention of information or formulation... extraneous cognitive load refers to the way information or tasks are presented to a learner; and germane... particular way of organizing and connecting information and a logic to the types and timing of questions...question constantly (and so should you!)  Amount of information and level of detail Again, this is an obvious...load, than certainly amount of attention and information input. If you’re reading a book, you only have...quickly they can understand and process your information. A class of three year olds is unlikely to be...I would think about how I was presenting the information. Maybe I needed to remove a distraction, such...

  9. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    Published
    Jan. 14, 2016, 3:30 p.m.
    ...copies of your predicted sequence information. Collect plasmid information in a binder or digital notebook...barcoded tubes and plates. The critical pieces of information for a given plasmid are the backbone, inserted..., the predicted full plasmid sequence. Other information may also be critical to the maintenance and use... with us, it will be easy to find all of the information needed for the process. Always sequence verify... Once the physical sample and its associated information are at Addgene, we are ready to sequence verify...always be derived from a single colony to ensure uniformity of the culture. Sequencing directly from the ...concentration, and not all providers are equipped to perform Sanger sequencing from glycerol stocks, so make...

  10. A Look at Addgene's QC Process

    Type
    Blog Post
    Published
    May 6, 2025, 1:15 p.m.
    ...confirmed or theoretical. No matter the level of information that is provided, each deposit goes through the...without the use of a reference sequence) using a bioinformatics pipeline developed at Addgene. The assembled...assembled plasmid sequence is then compared to the information provided by the depositor. If a depositor provides... will compare the NGS results to the plasmid information, other plasmids in the deposit, NCBI sequence... results and deposited sequences and plasmid information.  For each plasmid deposited at Addgene, QC scientists...scientist  The backbone elements based on the information provided and vector type  Any tags, selectable...sequence and assemble using the Illumina MiSeq platform, such as GC-rich regions and highly repetitive...

  11. Management for Scientists: Seeking Feedback

    Type
    Blog Post
    Published
    Jan. 8, 2015, 2:34 p.m.
    ...feedback in multiple formats      Ask for input and opinions in meetings, via email, at formal communications... hand and eagerness to get, as well as give, information to make improvements. Members connect directly... this means encouraging a culture of maximum information flow so everyone knows what is going on and can...break, go exploring outside the team, and bring information back. No team has all the answers – reinventing...frequently One of the most important paths of information flow is the one that brings feedback back to ...from at least some employees. Watch out for "information allies" – these are employees in your group that...communications events or at informal gatherings, like at lunch. Many experts recommend using anonymous surveys with...

  12. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    Published
    Oct. 4, 2018, 12:44 p.m.
    ...pooled libraries have allowed scientists to easily perform genome-wide screens to effectively and efficiently...sgRNAs per gene. These libraries provide more information while using less resources and are particularly...that improved sgRNA activity and thus library performance. The Root and Doench lab’s optimized genome-wide...CRISPRi, and CRISPRa libraries have been shown to outperform other popular CRISPR libraries (see below). Let...colon cancer) cells and assessed the libraries performance via the depletion of a gold standard set of genes...developed sgRNA-level metrics to assess library performance in viability screens. Based on these metrics ...sgRNAs, Brunello even with only a single sgRNA outperformed another CRISPR library with 6 sgRNAS, highlighting...

  13. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...repository that can be used to perform screens in human cells. General information regarding how to use a CRISPR...assembly cloning platform called GMAP (Gibson assembly-based modular assembly platform). GMAP is based ...-wide Screening Using CRISPR/Cas9”. Detailed information regarding the Wu Lab library can be found in ...orphan GPCRs demonstrating the utility of this platform for drug discovery. Researchers can use plasmids...stop codon) and closed (including a stop codon) formats. The collection is compatible with both standard...Gateway cloning and the FNLCR Combinatorial Cloning Platform (CCP); the latter of which greatly enhances the.... PubMed PMID: 26165496 GMAP - A new assembly platform for generating both simple and complex DNA constructs...

  14. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    Published
    July 11, 2017, 2:30 p.m.
    ...containing full information about the suggested gRNAs to use, including all information provided in the...phenotypic readout, similar mapping could be rapidly performed by targeting all potential components of a signaling...available through Addgene. This means you could perform a screen with one library from Addgene, and then.... Thus, it is always a good idea to keep a bioinformatician close if you need help putting your list together... in vitro, but there is enormous potential in performing in vivo screens, studying genes affecting cellular...several algorithms. Please read the “Detailed Information” text linked to each reference library before...adapters. Output_UserInputParams: Contains all the information that you have input in the web interface, making...

  15. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    Published
    Nov. 14, 2017, 2 p.m.
    ...found that many researchers feel that they lack formal training in a variety of transferable skills. At...many graduate and post-doctoral programs lack any formal or mandatory professional development training....you should have a business card; it’s a bit more informal, and a lot easier to hand out than a CV or resume...the connection you made at the job fair when you formally apply for the position. This post on Business ...call, make sure to get that person’s contact information and again, make a personal connection! Even if...are, and who they know all in one place! This information can be critical to a focused and effective job...probably have a lot of relevant experience and information to offer. However, if you hope to have a career...

  16. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ...protocols and performing rigorous quality control (QC). The specific QC experiments performed varies for ...of Pennsylvania Vector Core. Transfections are performed using the transfer plasmid, a plasmid encoding...and stored at -80 °C. Titer Titering is either performed by Addgene or by the University of Pennsylvania...Pennsylvania Vector Core. In general, titering is performed by the facility that produced the viral vector lot....Pennsylvania Vector Core is (as of April 2016) also performed by ddPCR.  Quality Control Addgene ensures high.... To learn which specific QC experiments were performed on your lot, please contact us. Full sequencing...the Viral Genome Next-generation sequencing is performed on viral genomes isolated from the final AAV preparation...

  17. Summer SciComm Series: A PhD in Science Communication

    Type
    Blog Post
    Published
    Aug. 23, 2022, 1:15 p.m.
    ...accepted as a grad student. I chose to focus on misinformation about sexual and reproductive health, as the...health issues creates conditions under which misinformation can flourish. I’ve never forgot about all the...perceive and evaluate peer-reviewed sources of information if those sources affirm or challenge their beliefs...term for a range of neat little tricks our minds perform to avoid cognitive dissonance (Kunda, 1990). More...conscious or unconsciously) pick apart sources of information they don’t like, looking for reasons to dismiss...scientific research is funded by the public and performed by researchers who received their educations from...schools and universities. However, much of the information gained from that research remains inaccessible...

  18. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...is, an antibody shown to perform well in one set of conditions may not perform as well in other conditions...assumed (Ayoubi et al., 2023). Additionally, their performance depends on the specific protocol used and the...For example, we have found that some antibodies perform well in a PFA-saponin fixation and permeabilization...population (Figure 1), further optimization can be performed to both maximize the separation and reduce any... is possible to design experiments where the performance of the antibody can be assessed through its ability...advantage of antibody validation efforts and information provided by external organizations like the Human...blood leukocytes. The file shared also contains information about the labeling pattern of several cell lines...

  19. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    Published
    May 24, 2018, 1:20 p.m.
    ...the students performing these experiments have a good baseline for comparison. Performing WISH using these...are too few resources to teach students how to perform these experiments in the beginning. In most classrooms...observation and dissections. Students don’t usually perform more modern techniques used in genetics or molecular...addition, the Sánchez lab has made it easier to perform advanced developmental biology experiments in planarians...both WISH and RNAi experiments, as well as how to perform worm amputations. They also created an online companion...RNA interference (RNAi) is a powerful tool for performing targeted knockdowns of genes of interest and ...mixing with the liver paste, these E. coli are transformed with the appropriate plasmid and dsRNA expression...

  20. Plasmid Preps: Different Purity, Different Quantities, Different Uses

    Type
    Blog Post
    Published
    June 16, 2020, 1:25 p.m.
    ...grade DNA Figure 1: Transformation introduces transformation grade plasmids into a bacterial cell...cell. Transformation grade DNA  Transformation is the process of inserting exogenous DNA in a microorganism...a plasmid preparation in 3 different ways: Transformation grade DNA Cloning grade DNA Transfection grade...plasmid DNA that is necessary for bacterial transformation of chemically and electrocompetent cells is...without commercially available kits and DNA for transformation doesn’t need to be as pure as for other uses...blog posts Learn about the differences between transformation, transduction, conjugation, and transfection...
Showing: 281 - 300 of 756 results