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We narrowed to 847 results for: PLE

Showing: 361 - 380 of 847 results

  1. Plasmids for Endogenous Gene Tagging in Human Cells

    Type
    Blog Post
    Published
    April 6, 2017, 1:02 p.m.
    ... features of the gene intact. Here, we show an example gene targeted for tagging with GFP at the C terminus...interest. For a C-terminal tag, as shown in the example above, we inserted a linker and GFP tag at the ...the GFP tag precisely where we need it, in this example, at the C-terminus of the gene of interest. To ...CRISPR/Cas9 (Cas9 protein and the crRNA-tracrRNA complex) and the donor plasmid into the cells we use electroporation... Institute for Cell Science Plasmid Collection, please visit the Cell Catalog on the Allen Cell Explorer...hiPS cell lines generated using these plasmids, please visit the Cell Catalog on the Allen Cell Explorer...

  2. Tips for Technical Support Calls

    Type
    Blog Post
    Published
    Sept. 20, 2016, 2:39 p.m.
    ...buried beneath someone's lunch. Many times, I’m pleasantly surprised to discover that the most crucial information...from you and it’s wrong. Can you help us fix it? Please address as soon as possible!” and I always begin...your concern is based on data you have produced, please share the data. Maybe not all, but many bio companies...helping us understand the nature of your concern. Please be courteous. We understand how exasperating it...time and money on a potentially faulty product. Please remember that the person at the other end of that..., let your customer service agent know (nicely, please!). We can’t improve without feedback. And if you...

  3. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    Published
    Dec. 1, 2015, 3:30 p.m.
    ...Having never been to China myself, I was pleased to be able to participate when the organizers of the...Lowe lab have allowed them to begin understanding complex regulation of and by tRNAs and tRNA-derived RNAs...long hypothesized that life started with RNA and a simple cell membrane. His recent work shows that they ...replication with no proteins). The chemistry is complex and this work requires an interdisciplinary team... they do it. Of course, No conference would be complete without CRISPR Michael Terns from of the University...structural and functional component of the ribosome complex and Dr. Steitz quipped that RNA in the ribosome...

  4. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    Published
    July 12, 2018, 1:09 p.m.
    ... passage number. Especially in a lab where multiple people are working on different things, this will ...in use. Wipe the hood with a disinfectant (most people use 70% ethanol, but you can also use Incidin wipes... this purpose.Learn about the effects of CO2 on sample pH Cell morphology: Shape and size matter You should...which multiplies every 16 hours). So if you, for example, split cells on a Monday diluting them 1:10, you...contaminated.Learn how Addgene uses barcodes to track samples Keep track of passage numbers Keep an eye on the...

  5. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    Published
    May 18, 2021, 1:15 p.m.
    ...to co-localize multiple enzymes in a reaction pathway, the MPyV system provides a simple, efficient method...SARS-CoV-2 by mutational scanning By Seth Kasowitz As multiple antibody-based vaccines are deployed around the...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...

  6. Tips to Make the Most of a Scientific Conference

    Type
    Blog Post
    Published
    May 23, 2019, 12:26 p.m.
    ...name for some context and maybe common ground. The people you meet may all be scientists but not all will...to practice beforehand. Generally, two types of people will visit your poster: those that want you to ...Try things like sitting at a table for lunch with people you don’t know or attending social events. I like...like to set goals for myself like meet ten new people each day. Exchange business cards. When you’re done...Follow up with those that you’ve met. Connect with people on LinkedIn or Twitter, for instance. If you’ve... head over to the “How to Make Friends and Meet People at a Scientific Conference” blog post. There’s ...

  7. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ...through homologous recombination. The concept is simple: deliver a template with a modified version of ...homology is typical for targeting constructs. In the example shown in Figure 1, large sequences corresponding...is expressed (knocked in). As seen in the above example, you can remove a floxed resistance gene using ...plasmids described in the preceding sections are simple ways of knocking out a non-essential gene of interest...there are 3 loxP sites, recombination can occur in multiple ways. The desired recombination event will remove...tools for creating knockout animals or cell lines, please consider sharing them by depositing with Addgene...

  8. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    Published
    Aug. 18, 2015, 2:30 p.m.
    ...strengthen your results. Enrichment or depletion of multiple gRNAs targeting the same gene can be strong evidence...jar of jellybeans, a pooled CRISPR library is a complex mixture. Pooled libraries are composed of many ...sorting and screened for a particular phenotype. For example, mutant cells can be used in drug screens to identify.... Loss of representation due to enrichment or depletion of specific gRNAs can lead to skewed results. ...any experiment, the use of appropriate controls, multiple replicates and several cell types can strengthen...

  9. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    Published
    May 29, 2014, 1:29 p.m.
    ...getting-started guide for BLAST, which includes a simple explanation of the different BLAST programs, databases...consistently run our blastx searches with the “Low complexity regions” filter unchecked so that these regions...benefit is that blastp searches do not filter low complexity regions by default, meaning that you do not have...Clustal is our frequent choice for pairwise or multiple sequence alignments of nucleotide or protein sequences... sequences. We also use COBALT for aligning multiple protein sequences, particularly for comparing different...//www.ebi.ac.uk/Tools/webservices/#multiple_sequence_alignment_msa http://www.ebi.ac.uk/Tools/webservices...

  10. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    Published
    May 20, 2014, 2:06 p.m.
    ...allowing very low background level of fluorescence. Examples for such photoactivatable or photoconvertible ... and their emission is green or red light. For example, T-Sapphire, LSSmOrange, and LSSmKate. Fluorescent...sensor), and iGluSnFr (glutamate sensor). More examples of these biosensors can be found at Addgene. ...also is also called Bimolecular Fluorescence Complementation (BiFC). 8 points to keep in mind when choosing...system can excite, and detect the emission. For example, if your microscope has only two lasers, at 488nm...minutes after it is translated to a few hours. For example, superfolder GFP (sfGFP) and mNeonGFP can fold ...

  11. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ... takes 1-1.5 hours to complete, while this syringe-based method can be completed in 20 to 30 minutes. ...drawbacks of kits and typically follow the same principles: cells are lysed, RNAs and proteins are removed... centrifugation. At this point, the plasmid DNA sample can be treated with RNase to remove RNA contamination...waste footprint. The Zhao and Deng labs developed a simple method for regenerating silica columns that come... reduce the number of spin columns you need to complete your research. Do you know of other nucleic acid...

  12. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

    Type
    Blog Post
    Published
    Dec. 3, 2019, 11:38 a.m.
    ...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...enough... If in the past you have tried to combine multiple terms in an Addgene plasmid search and found the...each filter is displayed in parentheses.  In this example I wanted to find a lentiviral plasmid to express...Starting with two search terms we see there are plenty of results, but I can’t browse through 300+ plasmids... plasmids that may be difficult to define for a simple search. These pages provide helpful information...

  13. Tips for arabidopsis transformation

    Type
    Blog Post
    Published
    Oct. 25, 2018, 1:23 p.m.
    ... of genes and proteins in living plants, to complementing mutant phenotypes. Arabidopsis is amenable to... sweet potatoes (Kyndt et al., 2015)! It sounds simple, and it pretty much is, but there are some tips...There are a bunch of great options, but here are a couple of my favorites. Plasmids from the Tsuyoshi Nakagawa...you transform early, your plants will still have plenty of time to produce seeds after you expose the flowers...for transformants. It’s very important to select multiple T1 plants from each transformation. I like to ... Agrobacterium T-DNAs with expressed genes: An example of a naturally transgenic food crop.’, Proceedings...

  14. Transferable Skills Guide: Identifying Your Transferable Skills

    Type
    Blog Post
    Published
    Jan. 11, 2018, 3:29 p.m.
    ...socks, persevered through a nerve wracking commute complete with a disabled train, and now you’re sitting ...within yourself, and to demonstrate them to the people who make hiring decisions. Though having all the...to other acquired skills.” It is nice when other people point out how awesome we are, but there is also...my wheelhouse”, take a second to reflect. For example, say your boss approaches your team with a project...front of a crowd and using your charisma to give people hope for some shiny vision on the horizon. You’...these responsibilities. If you are in charge of completing many meticulous protocols or tasks, you’ve probably...

  15. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    Published
    Aug. 9, 2016, 2:30 p.m.
    ...arranged in a double helix. People are wrong. In reality, cellular DNA is a complex three-dimensional globule...Technique What It Locates ChIP-seq Proteins (for our example later, histones) ChiRP-seq RNA Hi-C DNA (genome-to-genome...appendix DNA is a 3D fractal). Enhancer biology is a complex and dynamic field. We’re going to focus on a tried... beyondthedish (left) and biobabel (right). People commonly think of DNA as a linear polymer of A,...Bedtools getFasta. These regions and their reverse complements were parsed for spCas9 PAM sites (NGG) and then...

  16. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    Published
    Aug. 20, 2015, 2:30 p.m.
    ...expertise on the processes needed to successfully implement, manage and close-out a grant. Grant professionals...reports are submitted and project evaluations are completed. To enter and advance the profession, you can ...  Skills such as research, collaboration across multiple disciplines, writing, project management, and ...application writing and development, project implementation, program evaluation and grant management.  ... time and resources. 3. Get a mentor, find your people Find someone who is working or has worked as a ...knowledge of the 8 core competencies through a multiple choice exam.  The bottom line on being a grants...

  17. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    Published
    March 20, 2014, 3:04 p.m.
    ...to be managing. Being reminded of the seemingly simple actions we can take to help ourselves can be an...regular schedule at work. Choose a lab/job where people have regular schedules. Work 8 to 5 or 7 to 4 or...create “fake” family by forming a social circle of people in the same situation. Go to church, join a club... in the family to have to miss something – for example, say no to some of those birthday parties. Tell...print, but I actually let my daughter skip school a couple of times. Be sure to add time with your family,...time. For me, that’s been volunteer work at our temple, my kids’ school, and with professional organizations...

  18. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    Published
    Sept. 13, 2018, 1:55 p.m.
    ...al., guides were considered toxic if they were depleted from the screen. This toxicity was used a proxy... the growth screen, safe-targeting guides were depleted at greater rates than their non-targeting counterparts...Additionally, the distribution of enrichment and depletion scores for safe-targeting guides more closely ...noise, of dsDNA breaks caused by CRISPR. See supplemental figure 6 for more details. Safe-targeting guides...However, there was no significant difference in depletion of ricin sensitizer genes identified with short...Resources Watch Dr. Michael Bassik’s lecture on “Multiplexing with CRISPR Screens”    ...

  19. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    Published
    April 20, 2021, 2:30 p.m.
    ...mechanisms behind the cytotoxicity can vary. For example, the popular VSV-G envelope protein causes cells...are differentially inactivated by nonspecific complement mechanisms in serum. Sometimes this is due to...cells that the viral vectors were produced in. For example, production of vesicular stomatitis virus, HIV-...sugars end up in the envelope and are targets for complement-based killing by anti-alphaGal antibodies. The...good at infecting most cells, but not all. For example, LDLR expression in hematopoietic stem cells, T... as described above and is inactivated by the complement system in serum (DePolo et al., 2000). Other ...

  20. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...Its RNA-guided nature makes CRISPRa much simpler to implement in genome-wide screens over cDNA libraries... Rochester Medical Center. During development, complex genetic programs specify and assemble diverse arrays...individual genes can be relatively laborious because multiple plasmids/viruses are needed to deliver all the... then packaged into AAV for in vivo delivery. Multiplex gene targeting is also possible and is only constrained...vivo simultaneous transcriptional activation of multiple genes in the brain using CRISPR–dCas9-activator...
Showing: 361 - 380 of 847 results