Skip to main content

We narrowed to 928 results for: mpl

Showing: 421 - 440 of 928 results
  1. Click Biology: How to Click and Play to Explore Biology

    Type
    Blog Post
    ...achieve, but are simple to pick up and work well the first time. But biology is complicated — so many different...reactive groups of Click Chemistry are almost completely absent from living systems. What are the possibilities...functionality to existing genes, while being accessible simply by dialing up the right nucleic acid sequence (...homology-directed repair efficiency by linking the repair template VirD2 HUH-tag For creating DNA-protein conjugates...
  2. To Each HIS Own

    Type
    Blog Post
    ...proteins within a system. This can be pretty complicated given that a single cell has tens of thousands...strategy a single protein can be isolated from a complex mix using affinity chromatography against an amino...metal ions are immobilized on a bead or resin and a sample containing the protein of interest flowed through...interest and the metal ions.      Fig. 1: A sample containing a polyhistidine tagged protein of interest...tags are also recommended if you need a very pure sample. Because binding strength is proportional to the...
  3. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    ... that actually contain your insert of interest. Simply stated, ccdB makes cloning easier by selecting ...take up your insert. But exactly how does ccdB accomplish this? Let’s start with a brief history of the...system contains ccdB. A successful insertion will completely replace ccdB with the investigator’s insert of...interest.   CcdB resistant E. coli strains complete the system But ccdB alone isn’t the only key to...protein involves poisoning of DNA-topoisomerase II complexes.” J Mol Biol. 1992 Aug 5;226(3):735-45. PubMed...
  4. The Future of Research Symposium Boston 2015

    Type
    Blog Post
    ... about how the system operates and functions” “Complete lack of information on number of postdocs” “Postdocs...comments are well correlated with published data. For example, in 2010, less than 15% of postdocs had achieved...beginning to address these issues independently; for example, Harvard’s Therapeutics Graduate Program has a ...postdocs in academia, industry and other areas of employment? How many graduates and postdocs – and from what...organisations that represent postdocs and institutes that employ them, the diversity of trainee backgrounds, publishing...
  5. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ... the mutation or replacing the gene. A notable example is the treatment of Severe Combined Immune Deficiency...normally functioning T cell repertoire (1). In this example, it was possible to transduce the T-cells outside...based delivery techniques may hold the key. In the simplest form, translating gene therapies from conceptual...already have ways to prevent AAV infection. For example, prevalence of neutralizing antibodies against ...: 14566358.  2. Hirsch, M. L., et al. "Self-complementary AAV mediates gene targeting and enhances endonuclease...
  6. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ...CRISPR is a simple and versatile tool for genome engineering, but its utility is dependent on its ability... a fluorophore or tag. To utilize HDR, a donor template containing the desired sequence flank by homology...Nanoblades which carries a gRNA, Cas9, and a HDR donor template DNA can be generated by incubating Nanoblade particles...helps form DNA and retroviral-particle or VLPs complexes. The team found that "all-in-one" Nanoblades facilitated...facilitated HDR with either a ssDNA or dsDNA donor template. In vivo editing of mouse zygotes and mice with...
  7. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    ...coordination to more complex situations such as learning and memory or emotional states, for example anxiety or ...and cause variability in the testing group. For example, if an experimenter picks up a mouse from the home...kHz, produced from fluorescent light bulbs, for example) that are outside the humans’ audible spectrum.... have preferences for specific cell types. For example, AAV1 and AAV9 serotypes have strong neuronal tropism...
  8. How to Prepare for an Industry Interview

    Type
    Blog Post
    ...task, action, and results. While this method seems simple, I found it hard to remember to describe all four.... This is a chance to highlight your prior accomplishments and demonstrate your science communication ...interview. Expect to be tired once the interview is complete Industry interviews are long! Mine lasted 4.5 ...interviewer and then thank them once the interview is complete. Remember, your interviewers took time out of ...a question. Maybe you ask for questions after completing each section of your talk. Some video conferencing...
  9. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    ...process you would like to monitor? Neurons, for example, have unusually fast calcium dynamics, whereas ... is most sensitive to your expected range. For example, calcium in the ER lumen reaches the sub-millimolar...single fluorescent protein (FP), requiring only a simple measurement. However, single wavelength measurements...crucial advantage: tissue-specific targeting. By employing various AAV serotypes, researchers can precisely...Genetically encoded calcium indicators to probe complex brain circuit dynamics in vivo. Neuroscience Research...
  10. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...sequence can completely disrupt expression of the transcribed ncRNA (Jiang et al. 2014).  Simple knock-out...transcription of the ncRNA (Hazan and Bester 2021). For example, inserting a polyadenylation signal downstream ...interactions between ncRNAs and adjacent genes are complex and not yet fully understood, so it is important...variants may also be useful for targeting ncRNAs. For example, the Cas12a variant can cleave single-stranded ...
  11. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ... don't do anything. Viruses are complicated and they perform complex biological tasks, both of which ...virus-based experiment at one point or another. After amplifying a viral vector, collecting virus, and transducing...replication. To avoid genomic rearrangements, try amplifying the viral vector with bacteria designed to minimize...vector using the harvested viral solution as a PCR template. Regardless of the type of virus you’re working...
  12. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...and look at them under a microscope, fractionate samples to isolate particular organelles and their contents...fluorescent proteins have usurped old methods or complemented them in ways that make them much easier. A special...localized to a specific region of the cell. For example, if the ratio of red/green fluorescence falls within...protein or protein fusion. This one protein method simplifies cloning and makes your experimental setup less...in gene expression. In a developing embryo, for example, a researcher could determine whether or not newly...
  13. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...when studying them since their complete knockdown is lethal. Incomplete or titratable knockdown, however...the tools scientists can use to study them. For example, gene knockdown with CRISPR interference (CRISPRi...pathogenic strains and environmental isolates. For example, Peters and colleagues used Mobile-CRISPRi to knockdown...if that gene is a target of an antibiotic. For example, let’s take a look at the essential gene folA which...
  14. PhD Applications After COVID

    Type
    Blog Post
    ...know that this process only became even more complicated with COVID-19 and standards for hybrid, virtual... tends to go something like this:   However, as simply as it seems laid out above, there are many places...where the PhD application cycle becomes much more complex. “How do I reach out to potential mentors?” “Where...very valid questions, and often make an already complicated process even more difficult. I’ve learned that...potential programs and universities stressful or simply not feasible. Through the virtual systems created...
  15. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ...many aaRSs exist within a multi-tRNA synthetase complex, the team investigated whether the generated antibodies...antibodies were able to immunoprecipitate the entire complex in three different cell lines. The Gräslund lab...jbc.RA120.01289     Novel protein cage provides simple enzyme protection in yeast bioproduction pathways... a reaction pathway, the MPyV system provides a simple, efficient method for protecting a single enzyme...SaCas9. A new yeast inducible CRISPRi library - Ampl133 targets S. cerevisiae genes with 6-12 guides per...
  16. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    ...remove endonucleases (see figure at right for an example). Look out for carbohydrates Certain strains, including...the cell lysis process. To ensure that lysis is complete, use a reagent like Qiagen’s LyseBlue, which uses... uses color to show you if lysis is incomplete. Protein expression strains are not good for cloning Bacterial...yields. Why would volume make a difference? In the simplest of terms, more volume = more cells = more plasmid...the material used for inoculation. To further complicate matters, plasmid copy numbers are often ambiguous...
  17. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    ... plasmid page is nested in. You can see in the example above that the bread crumbs trail starts in the...publication. Addgene makes this easy by providing a template citation for you to use in your Materials and ... Use the Addgene catalog number to easily find template citations for Addgene plasmids you have used in...  These sequences are provided in FASTA format (example) and can be analyzed using Addgene’s Analyze Sequence...Price of course! The price of some items vary, for example, plasmid kits are avaliable for less than than ...
  18. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    ...incubated together without forming large oligomeric complexes. Antibodies are also too large to be expressed...primary antibody-nanobody mix can be added to your sample, thus eliminating the secondary antibody incubation...two-step staining with antibodies. Nanobodies also simplify multicolor staining experiments because they don...568, or Alexa Fluor 647 before application to the sample. Staining with this multi-color staining workflow...imaging, rapid epitope-mapping and native protein complex isolation. eLife. PubMed PMID: 26633879 PubMed ...
  19. Transferable Skills Guide: Identifying Your Transferable Skills

    Type
    Blog Post
    ...transferable skills you possess because any potential employers, collaborators, or co-workers want to know that...socks, persevered through a nerve wracking commute complete with a disabled train, and now you’re sitting ...sitting in the lobby of (hopefully) your future employer. Your heart is pounding, you’re short of breath, and...my wheelhouse”, take a second to reflect. For example, say your boss approaches your team with a project...these responsibilities. If you are in charge of completing many meticulous protocols or tasks, you’ve probably...
  20. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

    Type
    Blog Post
    ...including depositing labs and institutions. For example, search for “Melina Fan” and you’ll be able to ...plasmids from a paper have been deposited? In this example we’re going to be looking for recombinant monoclonal...each filter is displayed in parentheses.  In this example I wanted to find a lentiviral plasmid to express... filters you do not need to start a new search. Simply click the ‘X’ next to any filter to clear it. ... plasmids that may be difficult to define for a simple search. These pages provide helpful information...
Showing: 421 - 440 of 928 results