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Showing: 561 - 580 of 657 results
  1. The Challenges of Cell Culture

    Type
    Blog Post
    ...Henrietta Lacks. New York: Crown Publishers, 2010. 2. Gold, Michael. "A conspiracy of cells." State University...Marker-independent identification of glioma-initiating cells. The International Cell Line Authentication Committee...propagation of the line, and can include assays for authentication and contamination) and cell lines can become...let alone the human cost. Here are just a few publications which have since been retracted due to cell ...
  2. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ... PMID: 25184501. PubMed Central PMCID: MC4262738. 2. Doench, John G., et al. "Optimized sgRNA design to...is only effective when sgRNAs target a specific location on the gene. For CRISPRa, sgRNAs were targeted...Calabrese pooled library and lentiviral prep here! Applications for the Root and Doench CRISPR libraries Overall...et al. "A protein-tagging system for signal amplification in gene expression and fluorescence imaging....
  3. The Importance of a Fun Workplace: Company Culture at Addgene

    Type
    Blog Post
    ...interesting things to do, feel free to crowdsource! 2. Free food generally gets people to show up If you...be an obvious one, but don’t make things too complicated for yourself (especially if you are the sole ...sole event planner)! If you have a very intricate itinerary, there is a higher likelihood that something ...Delegate- it will save you time and sanity!    10. Communicate Make sure people know that there is something...
  4. Bioinformatics at Addgene

    Type
    Blog Post
    ...the necessary steps to perform:      Fig. 2: Quick-stepping to a FASTA file   With our outline... from depositing laboratories and associated publications, but our quality control (QC) standards have...such as 96-well microplates, and thanks to our dedicated product and development teams, frequent updates...can be easily shared with others without any complicated setup. We created, tested, and debugged each ...
  5. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...rate of 10-55% - the PASTE system.     Fig. 2: Schematic of programmable gene insertion with PASTE...experimental design and its potential for clinical applications. Specifically, efficacy rates of Cas7-11 were...place, while increasing the number of potential applications. But they weren’t done engineering quite yet... at the bench and potentially in therapeutic applications. The potential of CRISPR has yet to fully taken...
  6. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...GUS-staining is a very useful visual reporter (Figure 2). The GUS enzyme cleaves a substrate, which will then...vincristine, are regulated in the medicinal plant, the Catharanthus roseus (Figure 1). The ultimate goal of our ...and a vacuum is applied with a vacuum bell or desiccator. Successful infiltration can often be seen by...expression method for analyzing gene function in Catharanthus roseus seedlings." Frontiers in plant science...
  7. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    ...Hoogsten base pairs and wobble pairs.  Figure 2: (A) The codon wheel depicts the 64 codons that encodes...folding and stability, change post-translational modification sites and even affect protein function (Mauro... AP, Bilen B, Zavolan M (2013) Analysis of CDS-located miRNA target sites suggests that they can effectively...synonymous codon usage bias, and its potential applications. Nucl Acids Res 15:1281–1295 . https://doi.org...
  8. High Complexity Golden Gate Assembly with NEB’s DAD

    Type
    Blog Post
    ...rules are: (1) don’t use the same overhang twice; (2) avoid palindromes; (3) no overhangs with the same...assemblies are possible, the two sets of data together indicate that 35-pieces is a reasonable upper limit for...Overhang Ligation Fidelity by T4 DNA Ligase and Application to DNA Assembly. ACS Synthetic Biology. https...
  9. Transferable Skills Guide: Managing a Team

    Type
    Blog Post
    ...most important” tasks my schedule. This was bad for 2 reasons. First, it soon became obvious that I didn...delegating well, but it does get easier over time! Communicate often and clearly As a manager you need to clearly...it is the best way to avoid mistakes due to miscommunication and to detect trouble. I’m not comfortable...
  10. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    ...) or the American Chemical Society (ACS) are just 2 examples of organizations that hold large, annual ... conferences are held every year in different locations all over the world. But simply attending the conference...work. Addgene’s very own Joanne Kamens, a strong advocate for career planning, has written a great blog ...
  11. Plasmids 101: Codon usage bias

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    Blog Post
    ...PMID: 18478103. PubMed Central PMCID: PMC2364656. 2. Dittmar, Kimberly A., et al. "Selective charging ...ribosome may therefore stall at inappropriate locations or fail to make it through the entire transcript...generally segregate with the cell pellet during purification procedures. Even if you produce a large amount...
  12. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ...al (Nature. 1988 Jul 28;334(6180):320-5.) PubMed. 2. A segment of the 5' nontranslated region of encephalomyocarditis...desktop. The Plasmids 101 eBook is designed to educate all levels of scientists and plasmid lovers and...
  13. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    ...work. Nobody wins, and you need to make a change. #2 Thou shalt sometimes put work ahead of life Sometimes...other activities. Take care of yourself. Take a vacation alone with your partner – no kids. Get a massage...
  14. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...PubMed PMID: 3459152. PubMed Central PMCID: PMC323600. 2. Chester, Nicholas, and Daniel R. Marshak. "Dimethyl...desired modification. Finally, the positive clones are sequenced to confirm the desired modification and ...polymerase activity (for amplification), 3’->5’ exonuclease activity (increases amplification fidelity), and no... genome modifications. Occasionally, multimerization of the PCR primers can cause duplication of the primer...the desired modification have been identified (and confirmed to be free of primer duplications), final validation...and the absense of additional modifications.   Site-directed mutagenesis of Plasmids. A) Generation..., which flank the targeted region. Upon PCR amplification, the template is removed by DpnI restriction...
  15. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...gene1 [lon] ompT gal sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R(zgb-210::Tn10--TetS) General protein expression...expression vector cannot have p15A origin of replication F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) pLysS...expression vector cannot have p15A origin of replication F- ompT lon hsdSB (rB- mB-) gal dcm (DE3) pLysE...Expression vector cannot have p15A origin of replication F- ompT hsdSB(rB- mB-) gal dcm (DE3) pRARE2 (...Expression vector cannot have p15A origin of replication fhuA2 [lon] ompT gal (λ DE3) [dcm] ∆hsdS/ pLemo...Expression vector cannot have p15A origin of replication F-, Φ80ΔlacM15, thi, lac-, mtl-, recA+, KmR Expression... commercial strains carry the DE3 lysogen, as indicated by the name of the strain. Conversely, other strains...
  16. Transferable Skills Guide: Creativity

    Type
    Blog Post
    ...to the break room for a sip of water.  Fig 2: Blugene likes to spark creativity in out on the open...resources you’ll need ahead of time and mute your notifications during. (Boyes, 2019) If you’ve taken the time... particularly relevant to you. Elect a devil’s advocate. This is the person whose job it is to disagree...think about alternative options. When a devil’s advocate is elected, others may feel they can share opposing...dissenting opinions or ideas. While the best devil’s advocate is one who truly disagrees with the group (because...adamantly for their solution), selecting a devil’s advocate is a good option for any meeting in which the ...other way,” you probably have reason to continue advocating for your idea. Our product team has implemented...
  17. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ... PMID: 9537413. PubMed Central PMCID: PMC3010411. 2. Kohli, Manu, et al. "Facile methods for generating...gene at once. Therefore, applications that require simultaneous modification of all alleles of a gene,... recommends designing homology arms with the modification of interest, ordering them from a gene synthesis...with a homologous repair template. It is this application - the creation of sequence-specific knockins ...clones, making it unsuitable for most in vivo applications.  Third, even in the most optimized tissue-culture...tissue culture based approaches that require modification of only a single allele of a gene, such as the...efficiently deliver large repair templates for applications that require large gene fusions. The current...
  18. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...of 3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a fluorophore modified from DsRed. DFHO...aptamer soup by exploring the development and applications of fluorescent RNA aptamers in Addgene’s collection...target cellular RNA to specific intracellular locations using Cas9 and track the localization of the complex...biotin (the addition of biotin allows for the purification of specific RNAs from cells using this tag, ...TO1-biotin and TO3-biotin contain an acetate modification that reduces nonspecific binding with RNA by...non-coding RNAs in mammalian cells." Nature communications 9.1 (2018): 656. PubMed PMID: 29440634. PubMed...fluorescent rna aptamer spinach2 for imaging of replication and immune-mediated changes in intracellular ...
  19. Thoughts on the Future of Research 2015 Boston Symposium

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    Blog Post
    ...Postdoctoral Associations around Boston - From Panel Session 2: Defining the Postdoc: An Institutional Perspective...the FOR website, the FOR Boston 2015 organizers indicate that the main question they wanted to focus on...oriented, scientifically minded, and able to communicate complex information - all traits you are likely...Publishing Panel brought in editors from a variety of publications as well as young PIs and all gave advice on ...blogging, write reviews, or write for a university publication, improving your general writing skills will help...academic publishing goals and make you a better communicator overall. Don’t Write in a Vacuum Your research...cognizant of this during my PhD. Submitting a publication doesn’t have to be like throwing it into a black...
  20. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...Nature 339 (1989). 487–488. PubMed PMID: 2725683. 2. Miller, C.J., Kassem H.S., Pepper S.D., Hey Y., Ward...positively charged polycation, polybrene. Interestingly, one study found that alternative polycations such as DEAE-dextran... freshly collected lentivirus for downstream applications. While this ensures that the highest titer virus...lentivirus may want to consider testing a variety of cation and serum sources to ensure the best possible transduction...establishes the best conditions for specific applications, stick with them! Changes in such variables ...efficiency of lentiviral vectors: effect of sera and polycations.” Molecular Biotechnology 53.3 (2013). 308-14...
Showing: 561 - 580 of 657 results