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We narrowed to 1,009 results for: PLE

Showing: 181 - 200 of 1009 results

  1. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    ...expediently join multiple synthetic fragments, create multiple mutations in one or multiple fragments, and...joining multiple fragments of DNA to create a synthetically designed DNA sequence. There are multiple methods...becomes as simple as mixing fragments together with linearized vector for seamless joining of multiple mutations...standardized constructs and tools for the rational and simple shuffling of DNA regions. Golden Gate Assembly ...Assembly Tool which is freely available online. Its simple interface can guide you through both experimental...Assembly Mix include:  Generation of constructs with multiple point mutations Cloning sgRNAs Production of Yeast...Expression Cassettes Generating constructs with multiple point mutations NEBuilder HiFi can simplify multi-position...

  2. Networking Follow-up: I Met a Person. Now What?

    Type
    Blog Post
    ...important” or senior people. Focus on building relationships with peers and people around you naturally...run. Don’t worry about the people that don’t respond; focus on the nice people who do respond. Don’t call...connecting with people you don’t like or who do not show interest in you. Network with the people that you want... it’s about building genuine relationships with people you care about knowing. How do you follow up with...the encounter. As a side note, my guess is that people will not be exchanging business cards for awhile... you have something to ask or say. It can be as simple as “it was nice meeting you.” Include follow-up...who helped you succeed in any way. Don’t believe people who tell you that thank you isn’t necessary. I ...

  3. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...and so it may be necessary to try multiple techniques. For example, we have found that some antibodies...detecting the target-of-interest in your assay and sample-of-interest. The approaches available broadly map...determining the selectivity of antibodies for other sample types and for non-surface (intracellular) antigens...protocol used and the abundance of the epitope in the sample, relative to cross-reactive antigens – that is,...approaches used will be different depending on the sample and target type. To start the validation process...antibody validation, and it is recommended to screen multiple candidate antibodies when possible. If cost is...the target at a robust level, similar to that in samples of interest. If you are working with cells that...

  4. Developing Lab Management Software for Biology

    Type
    Blog Post
    ...success or failure: In our second example we’ve added some simple but critical logic. Some steps fail... a new tube. In the first example we’ve used inheritance to tightly couple the contents of a tube with...appropriate glycerol stock within our freezers Pick the sample into the appropriate stab culture Grow the stab...proper location To keep ourselves from mixing up samples and to ensure that everything is grown successfully...needs to handle these cases. The following code examples show how software can be structured to allow for... for the messiness of biology. These examples are intentionally trivial, but they're here to help tell...tell this story. In this first example to the right (# this will make you sad),  someone has put in a request...

  5. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...compatible with multiple FP(11) fragments, even from a different base FP (for example, EBFP2(1-10) can...generally known as Bimolecular Fluorescence Complementation (BiFC). It can provide a qualitative or quantitative... or even single-particle tracking of the bound complex. There are so many FPs available, with many ways...because they are used with an already-available complementary fragment. Next, let's look at a round-up of...EBFP2(1-10) and Capri(1-10) for use with GFP(11) Multiplexed labeling of cellular proteins with split fluorescent...Cerulean Cerulean(1-10) for use with GFP(11) Multiplexed labeling of cellular proteins with split fluorescent...protein fragments for bimolecular fluorescence complementation analysis under physiological conditions. Shyu...

  6. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...DNA strands complementary and non-complementary to the gRNA, respectively. When complexed with a truncated...full-length gRNA (20nt complementarity to target site) or truncated gRNA (≤15nt complementarity to target site...with which it is complexed rather than the protein itself. When native Cas9 is complexed with a gRNA that...transcriptional activator to the Cas9-gRNA complex. When this complex is directed to a target upstream of the...bioengineering, as it has facilitated the development of a simple and scalable means of making targeted genetic edits... directed to virtually any genetic locus when complexed with an appropriately designed small RNA, or guide...conventionally contains a 20-nucleotide sequence that is complementary to the target site, or protospacer, in the genome...

  7. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    ...group and topic. For example, it may be useful to adapt the second version (multiple presenter, figure by...time that accommodates more people. The feedback could be solicited via multiple avenues including casual...in journal clubs. They also have likely had the pleasure of attending both productive and not so productive...and maintain. It became clear that while several people across teams had various levels of experience with...decisions to make. How often will you hold it? How many people do you expect to attend? What is the expectation...ensure success. One of the most important is letting people know it’s happening. Aim for 3-4 weeks advance ...advance notice for the first meeting so that people can set aside time in their schedules and read the paper...

  8. Summer SciComm Series: Modes of Communication

    Type
    Blog Post
    ... pros of multiple channels/balance out cons. Flexible. Can allow people to learn in multiple ways from... means. Easy to combine multiple types of media. High barrier to entry (people have to enter a building... often unintentionally plays a big role in what people think of as "the best way" to communicate their...work just as well for everyone else?  In truth, people are incredibly varied in how they prefer to consume...However you choose to communicate your science, some people will love it and others will avoid it like the ...socioeconomic status and geographic location. For example, not all social media platforms are available or...probably didn't sound like a great idea to most people in 2005, but XKCD is pretty successful!  What resources...

  9. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ...by expressing multiple crRNAs in parallel (Fig. 3), multiplexed insertions into multiple distinct targets... The requirement for multiple Cas proteins to form the Cascade targeting complex is a hallmark of Type...nuclease activity. Instead, it forms a novel co-complex with the transposition protein TniQ for targeting... by the system. Guided by the crRNA, QCascade complexes survey the genome and bind target DNA sequences...pairing. Separately, TnsABC forms a transposase complex with the mini-transposon, excises it from the donor... report of VchINT, three separate plasmids and multiple promoters were used to express the necessary components...in single-plasmid INTEGRATE (pSPIN), which offer simple transformations for smaller cargos, and the ability...

  10. Hot Plasmids - May 2022

    Type
    Blog Post
    ...barcodes for live cell clonal multiplexing by: Rachel Leeson Multiplexed images in live cells remains ...own subclone, they were able to develop a 12-plex multiplex imaging system. Mixing the subclones within...effects of hi-multiplexing within in a cell, while potentially allowing for up to 20-plex experiments with...ever like to write about a recent plasmid deposit please sign up here. Here's what you'll find in this...knock-ins Visual barcodes for live cell clonal multiplexing Improving flexibility for in vivo proximity...Viviana Gradinaru Lab has previously developed the multiplexed version of the Cre-mediated AAV Targeted Evolution...mice and nonhuman primates (for AAV.CAP-B10, for example).  Given that the liver is an immunologically active...

  11. Advice for Starting a Biotech Company from a Grad Student turned Entrepreneur

    Type
    Blog Post
    ... into Sample6 as graduate students in Dr. Jim Collin’s Lab in 2009. From 2009 to today, Sample6 has changed...leadership, and grown to 25 members.  What is Sample6? Sample6 is an enrichment-free bacterial pathogen detection...their idea at various levels of complexity and abstraction and convince people with diverse backgrounds how...startup company. However, like many like-minded people, I quickly realize that a company requires more...current VP of Business Development & Operations of Sample6, for advice on how to take an idea developed during...etc. What project in the Collins lab inspired Sample6? The initial work focused on using bacteriophages... the focus of the application was changed, but Sample6 still leverages the core engineering and scientific...

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

    Type
    Blog Post
    ...colleagues had moved on as well, so I knew very few people. I didn’t really fit in anywhere and everyone who...the meeting. I didn’t know anything about these people at the time, but I met some very accomplished scientists...with theirs.  It was at this dinner and the next couple of days of discussion with my dinner partners Denise...would provide a structure and support network for people attending large meetings. A key was to provide ...bin with a mentor who had attended the meeting multiple times before and would help the Singletons plan...help with the program on social media. Many of the people who participated as mentors or promoters were connected...vastly expand their networks as they naturally find people with which they have interests and experiences ...

  13. Transferable Skills Guide: Teamwork

    Type
    Blog Post
    ...help of these people and others, we have updated our kit process substantially and have plenty of ideas for...group of people with distinct skills who fulfill a long-term function. When a select group of people are responsible...with other people, whether you want to acknowledge that or not. I just Googled the phrase “People need social... a leader can be successfully doled out to multiple people: someone to coordinate, someone to decide, ... have a specific purpose and finite goals. For example, when it came time to update how we make and distribute...function. Project team A Project Team is assembled to complete a series of tasks with precisely defined goals... in which case whoa! Mission accomplished! One example of a mission-based team in academia would be your...

  14. Intro to Our Summer SciComm Series!

    Type
    Blog Post
    ...the process of communicating science to laypeople.  Laypeople are simply non-experts in the field of science...would all be defined as laypeople when it comes to molecular biology.  Some lay people have a high amount ...other uses. Many laypeople would use “PCR test” to mean “a test that uses a biological sample to look for ...Table 1: Examples of defining your audience for science communication activities  Many people are concerned...language is aimed at a peer audience.  Note that lay people can and do consume scientific communication (it...use-case (definition) in a specific field.  For example, while “PCR test” has recently transitioned from...the possibilities are endless.    Activity Example audience  Talking about your career as a scientist...

  15. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    ...), which allows for the following to occur: A) Multiple polyclonal secondary IgGs can bind to different...sites per antibody. C) IgG’s are big enough that multiple labels (green circles in figure 2 below) can be...incubated together without forming large oligomeric complexes. Antibodies are also too large to be expressed...Antibodies Bivalent Polyclonal Can be labeled with multiple dyes Commercially available Not recombinant...sacrifice Good tissue penetration Can be labeled with multiple dyes Greater resolution due to lower label displacement...strength to a standard IgG secondary antibody (Pleiner et al figures 4A-C). And since they have limited...for colocalization labeling of up to 3 targets (Pleiner et al figure 4D). Unlike large secondary antibodies...

  16. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...BioRender.com.   Opto-G-protein coupled receptors  G-protein coupled receptors (GPCR) regulate cellular...consider when choosing one.     Figure 1: Example of an experimental optogenetic setup. A light source...light-gated ion channels, optogenetic G-protein coupled receptors (OptoXRs), and photoswitches. Light-gated...within a narrower band of wavelengths to allow multiple tools, such as biosensors, to be used in tandem...cellular function through multiple intra-cellular signaling cascades. Optical methods for controlling GPCR...than traditional pharmacology techniques allow. Multiple photoswitches can be used in a single pathway ...awry in a disease state.      Figure 4: Example of an intracellular signaling pathway activated...

  17. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ... type of multimer present in your sample. In these instances, please note that the supercoiled dimer band...plasmid monomer from a plasmid sample, or if no monomers exist in your sample, you could try linearizing ...known as multimers, and they are the result of multiple plasmid copies combining in a process known as...control when performing a diagnostic digest is a simple but effective method for detecting plasmid multimers...Bedbrook & Ausubel, 1976). In fact, if your plasmid sample has a mix of different-sized multimers, you may...agarose gel). If the presence of a multimer in your sample remains unclear after visualizing it on a gel, ...consistent with the size of a supercoiled plasmid dimer. Please note that it is common to see other plasmid confirmations...

  18. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    ... in-frame protein fusions, and multiple cloning sites. In the simplest case we have a standard backbone... that consistently receives around 200 new DNA samples every week. Here we will provide an inside look...own plasmids. Staying organized With hundreds of samples coming through our doors, the first step to ensuring...our deposit process, and then we carefully track samples through our lab using barcoded tubes and plates...growth instructions (temperature, strain, media supplements), suggested sequencing primers, or mammalian ...assemble your expected full plasmid sequences (examples include Benchling, Serial Cloner, and Snapgene...process. Always sequence verify Once the physical sample and its associated information are at Addgene, ...

  19. Working with Nuclear Receptors

    Type
    Blog Post
    ...coregulators exist as multiprotein complexes, a member of a single coactivator complex can serve as a rate-limiting...conduit to control the actions of the whole complex.  For example, the phosphorylation status of SRC-3 defines...afforded by coactivator complexes suggests that coactivators may be integrators of multiple cell signaling systems...datasets, for example, document the expression patterns of nuclear receptors across multiple different tissues...interact directly with NRs) exist in steady-state complexes with secondary coregulator (or co-coregulator)...co-coregulator) partners. The coregulator ‘core’ complex is composed of a tightly-bound invariant group of proteins... bound co-coregulators associate with the core complex in a dynamic, regulated manner. The fact that coregulators...

  20. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    ...result is a heteroduplex, with overlapping strands of edited and unedited DNA. The heteroduplex is resolved...any base-to-base conversion in principle, but some changes require multiple steps and enzymes. Prime editing...repair allow for correct heteroduplex resolution? The prime editing heteroduplex is likely recognized as...CRISPR guide. Some examples include: TwinPE: two prime editors make complementary edits to opposite strands...editing is also more precise. Base editors, for example, typically edit all the target bases within the...edited sequence on the other strand. To guide heteroduplex resolution to favor the edit, the Liu lab turned...component of the MutSα–MutLα mismatch repair (MMR) complex, to temporarily inhibit this process and promote...
Showing: 181 - 200 of 1009 results