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We narrowed to 847 results for: ple

Showing: 301 - 320 of 847 results

  1. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    Published
    Oct. 13, 2022, 1:45 p.m.
    ... relevant bases within the editing window, for example all C’s will edit to T’s. Thus, it’s important ...knocking out a particular gene isn’t feasible. For example, temporary repression or knockdown allow you to... target genes using dCas9-based activators The simplest of the dCas9 activators consists of dCas9 fused...SunTag system which facilitates recruitment of multiple activation domains to the same genetic locus via... proteins. The Synergistic Activation Mediator complex, generally the strongest activator, consists of...Edit”. Is Cas9 not your Cas of choice?  There are plenty of other enzymes to choose from within the Cas ... Multiple Authors...

  2. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    Published
    April 13, 2021, 1:15 p.m.
    .... The optimized plasmid curing method was much simpler and took only ~32 hours instead of ~60 hours. ...strategy for plasmid curing, providing users with a complete editing process. The MUCICAT system While the ...including concatenations into crRNA arrays targeting multiple different sites in E. coli. pCutamp cures the ...Ptet, and Para. The pDonor plasmid pRE57-Ter with multiple terminators can serve to interrupt genes (or gene... pQCascade) with 4 rounds of transformation to complete the N-acetylglucosamine strain library construction...dosages to facilitate swift optimization of strains. Multiple induction systems to choose from. Ready for metabolic...mainly on the development and application of a multiplex genome editing system based on CRISPR-associated...

  3. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    Published
    Dec. 14, 2017, 2:08 p.m.
    ...blood sample, in the best-case scenario [7]. In the clinical setting where patient blood samples are limited...especially if used to make clinical grade hiPSCs. For example, the CytoTune™-iPS 2.1 Sendai Reprogramming Kit...persistence of residual viral material [6]. The complete removal of the residual viral particles and establishment... in the presence of these two factors are not completely understood, the addition of an shRNA for P53 ...grade hiPSCs from easily accessible patient blood samples. All pCXLE vectors listed below in Table 1 are ...cells from CD34+ cells across blood drawn from multiple donors with non-integrating episomal vectors. ...

  4. Career Insights: Technical Support Specialist

    Type
    Blog Post
    Published
    Nov. 9, 2017, 2 p.m.
    ...the progress of the project. After that rather unpleasant experience I thought: I could do this job so ...companies produce different products and therefore need people with different areas of expertise in technical .... In addition, it can be advantageous to speak multiple languages - that makes communication with customers...are different names for technical support roles. Examples are Technical Service Representative, Technical...optimize” a particular standard lab protocol. For example, someone may attempt super quick incubation times...product in question. This can solve the problem in a simple way even if you don’t exactly know why. Personal... use these communication skills by working with people from many different backgrounds and realizing that...

  5. Healthcare Consulting: A Door to the Business of Life Sciences

    Type
    Blog Post
    Published
    Oct. 13, 2016, 2:30 p.m.
    ...consultants who enjoy their jobs most, get pleasure from taking up such complex challenges, quickly breaking them...got my PhD a couple of years ago in Bioengineering, and one of the things that pleasantly surprised me...Healthcare consulting is a fast-paced field, requiring people who are willing to quickly learn, and apply their...The types of projects mentioned above are just examples, and healthcare consulting firms can be involved... career in several different fields. While some people continue to enjoy the challenges of healthcare ...

  6. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    Published
    March 12, 2019, 1:37 p.m.
    ...chalk talk? On the surface, a chalk talk is quite simple. It's you, a dry erase board, some markers (chalk...question, not a vague goal or a "what" question. For example, "we want a better understanding of the Z pathway...acceptable. Aims should be mechanistic, doable by 1-3 people in 1-5 years, and contribute to resolving the overarching...scientists outside your field, and interesting. For example, “there is a significant need for healthy livers...explain why it’s significant and innovative. For example, "while it is possible to grow liver stem cells...chalk talk Perfect your story and delivery. Design simple figures and diagrams to illustrate your vision....also helpful to practice at least a few times with people that are outside your lab and outside your field...

  7. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    Published
    Nov. 1, 2018, 12:35 p.m.
    ...designated this fragment as the ccdB region (for “coupled cell division”). The encoded toxin-antitoxin system...subunit of DNA gyrase and trapping it in the cleaved complex. This results in DNA breakage and cell death. Conversely... so that the marker can be reused to introduce multiple genetic modifications in the same strain. MazF...translation. Here, Mok and Hok translation is coupled. Translation from the Shine-Dalgarno (SD) sequence...often lost without plasmid addiction systems. For example, Cara Morin and James Kaper used the hok/sok system...Teru, and Sota Hiraga. "Mini-F plasmid genes that couple host cell division to plasmid proliferation." Proceedings...K. Burley. "Crystal structure of the MazE/MazF complex: molecular bases of antidote-toxin recognition....

  8. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    Published
    Aug. 13, 2019, 12:58 p.m.
    ...whole-genome/transcriptome” studies, in which, for example, the activity of all genes in response to an experimental...the globe (Nüsslein-Volhard et al., 2012). One example (which I will come back to) is the one-eyed pinhead...morpholinos. When injected into zygotes, morpholinos complementary to a target mRNA inhibit its translation either...genome is useful for a variety of applications. For example, specific tissues such as blood cells or neurons...regulatory systems have also been successfully implemented in zebrafish. Developmental biology insights ...study profiled 38,731 cells from 694 embryos over multiple stages and reconstructed the patterns of gene ...well-suited for neuroscience investigations. For example, neuronal activity can be tracked in whole brains...

  9. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    Published
    July 25, 2019, 12:03 p.m.
    ...surface) of the leaf. This is a simple and easy technique to implement. Success might vary between experimenters...tobacco, Agrobacterium infiltration into leaves is a simple and efficient way to overexpress a gene and evaluate...your experiment (de Ruijter et al., 2003). For example, we did our initial transformations with the GUS... on transformation success. Some reporter gene examples are below. The GUS gene (β-glucuronidase) with...temporal reporters to measure promoter activity, for example. Useful positive controls could include: pSB96 ...expression from Agrobacterium, but not the plant. For example, we have seen strong expression of the GUS gene...

  10. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ...coming weeks, but, in this post, we will outline a simple method for selecting the guide RNA, validating ...CRISPR Design tool supplied by the Zhang lab. This simple website invites you to input the target region ...happy with your selection, ordering the sgRNA is simple and easy. Several companies supply gRNA sequences...you gRNA in vitro first. In vitro screening is a simple and fast method for validating your selected gRNA...). Testing the efficacy of your gRNAs is then a simple matter of transfecting HEK293T cells with your ...perfect primer sets to use to genotype your pups. A simple PCR with these primers will let you know if there...-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated genome engineering...

  11. The Challenges of Cell Culture

    Type
    Blog Post
    Published
    May 5, 2016, 2:30 p.m.
    ...put, the majority of cell line problems come from people sharing cells: the black market of cell biology...when it is important to check their identity. For example, perhaps a working cell bank has been made by a...stocks of cells. Working stocks should be frozen in multiple vials and thawed to produce a new batch once you...the cells you are working with because you have plenty of maintained freezer stocks. It is vital that ...monitor any changes. Your tracking system could be as simple as a piece of paper on the front of the incubator...passage number, cell line, and date, which you complete each time you passage the cells (as well as recording...are changing but yet more is needed to convince people of some of the basic tenets of responsible cell...

  12. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    Published
    April 7, 2016, 2:30 p.m.
    ... for using lentivirus, infect and select, seems simple, in actuality, many find using lentivirus to be...al. outline a do it yourself PCR-based approach complete with freely available positive controls. Routinely...labs prefer to collect and pool the harvests from multiple time points while others opt for a single harvest...transfer and packaging plasmids. A benefit of the multiple harvest strategy is that each day you are supplying...destabilize the viral envelope. One drawback of the multiple harvest approach is that it tends to result in...buffers used, and other experimental parameters. For example, while freeze thaw is generally not recommended...that lentivirus produced at pH 6 are resistant to multiple freeze thaw cycles. Consequently, it is best to...

  13. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ...proteins. lon protease: Strains where this is completely deleted (designated lon or Δlon) similary reduce...eukaryotic proteins Lemo21 (DE3)* Chloramphenicol (pLemo) Rhamnose-tunable T7 RNAP expression alleviates...replication fhuA2 [lon] ompT gal (λ DE3) [dcm] ∆hsdS/ pLemo (CamR) Expression of toxic, insoluble, or membrane...variety of sources, so fear not—there are a few simple troubleshooting measures that can help get you ...will not express in an IPTG induction strain for example, nor will a p15 plasmid be compatible with a pLys...cell-density they support; however, minimal media supplemented with M9 salts may be preferable if the protein... Proteins which contain hydrophobic regions or multiple disulfide bonds may aggregate and become insoluble...

  14. Designer PUF Proteins for Any RNA Target

    Type
    Blog Post
    Published
    June 17, 2014, 7:57 p.m.
    ...a world of possibilities in the engineering of complex gene networks. A comparable system for targeting... of RNA is highly desirable for extending the complexity of genetic circuits, allowing for tighter spatio-temporal... of interest. The new tool was developed and implemented by applying the Golden Gate cloning method to...March is fused to a translational repressor. In principle, however, designer PUF proteins can be fused to...in addition to their use in basic biology, for example, by targeting pathways underlying cancer or other...Assembly Kit out for themselves, she does have a couple of tips: First, she recommends using standard BsaI...

  15. Quick Guide to Near-Infrared Fluorescent Proteins

    Type
    Blog Post
    Published
    Oct. 5, 2021, 1:15 p.m.
    ...Institute for Biological Studies) Multiple uses: From multiplexing to multiscaling NIR-shifted spectrum...spectrum is perfect for use in spectral multiplexing (Shcherbakova et al., 2018). NIR FPs can be combined ...Verkhusha, V.V. Near-Infrared Fluorescent Proteins: Multiplexing and Optogenetics across Scales. Trends Biotechnol...11, 239 (2020). Oliinyk, O.S., Shemetov, A.A., Pletnev, S., Shcherbakova, D.M. & Verkhusha, V.V. Smallest...cyanobacteriochrome as versatile tag for spectral multiplexing. Nat Commun 10, 279 (2019). Oliinyk, O.S. et...Huisman, T.M., Verkhusha, V.V. & Hodgson, L. Direct multiplex imaging and optogenetics of Rho GTPases enabled...

  16. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    Published
    Jan. 29, 2019, 2:27 p.m.
    ...locations using Cas9 and track the localization of the complex. Broccoli, the other green fluorescent aptamer,... RNA. Fluorescence is observed when these RNAs complex and the entire aptamer can hybridize and fold. ...uniqueness of the photostability of the Corn-DFHO complex. For dessert (last but not least), the only fruit-named...fruit-named aptamer, Mango, binds to multiple derivatives of thiazole orange (TO) and fluoresces in the...Other groups have used Mango to study RNA-protein complexes (Panchapakesan et al., 2017) and short non-coding...mango aptamer-fluorophore: a bright, high-affinity complex for RNA labeling and tracking." ACS chemical biology...transcription using a photostable RNA–fluorophore complex." Nature chemical biology 13.11 (2017): 1187. PubMed...

  17. 10 Steps to a Perfect Science Talk

    Type
    Blog Post
    Published
    Aug. 23, 2016, 2:30 p.m.
    ...resources on self-promotion (how hard it is for some people, especially women), visibility (how to get it, ...especially if introverted ), and networking (how to get people to remember you). What better way to accomplish...Interviews: The All-Important Job Talk. 1. Watch other people give great talks We are not talking about lab ...simplicity of their slides and how the slides complement the spoken message. Learn from their level of...cause and effect. Stories can and should be very simple. The goal is to get the listeners to be thinking...on a slide but not reading the explanation. The people that need the information will find it. Experienced...particular question. I like to ask about how many people use plasmids in their research and then see how...

  18. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ...Brainbow multicolor labeling system - a perfect example of the continued utility of Cre-lox. Check out ...events specified by three loxP variants, but the principles remain the same. The first fluorescent protein...than 3-4 colors due to the tandem integration of multiple copies of the construct (about 8 in Brainbow-1.0...-1.0 mice.) The combinatorial effect of multiple, independent recombination events leads to a rainbow ...Brainbow-2 each contained a “default” state; for example, Brainbow-1.0 expresses RFP when the construct ...system, Sanes and Lichtman also developed a complementary Flpbow construct that is functionally similar...13):2835-46. doi: 10.1242/dev.094631. PubMed. Multiplex cell and lineage tracking with combinatorial labels...

  19. Searchable and Sortable gRNAs for Your Next CRISPR Experiment

    Type
    Blog Post
    Published
    May 17, 2016, 2:30 p.m.
    ... by multiple keywords that are separated by a single space to get even more specific. For example, to ...e.g. SNPs) exist. Additionally, consider testing multiple gRNAs targeted to the same locus to ensure their... are not generated from a plasmid. If you, for example, prefer to deliver your CRISPR components as RNPs...and you can search by any part of the data. For example, entering ‘C. elegans’, or ‘elegans’ will filter...publication. As you develop and confirm new gRNAs, please consider submitting their sequences (and plasmids...

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

    Type
    Blog Post
    Published
    July 21, 2015, 2:30 p.m.
    ...DePace’s yearly planning meetings revolve around two simple worksheets, each of which is filled out separately...summarized briefly below (and available of in the supplement of this paper), help each person organize his...including major project goals and effort needed to complete them. These documents dictate the subject matter...concrete plan in hand, trainees are motivated to complete their work rather than overwhelmed or confused...meetings may not look the same in every lab, but the principles behind this technique are beneficial for researchers...Extracurricular Activities for a Strong Science Career Path 7 Simple Strategies to Resolve Conflict With Difficult Supervisors...
Showing: 301 - 320 of 847 results