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We narrowed to 704 results for: ANG

Showing: 81 - 100 of 704 results

  1. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    Published
    Aug. 5, 2025, 1:15 p.m.
    ...Other PCR conditions can be changed to further lower fidelity, like changing the metal cofactor and the...This post was written by Alyssa Shepard and Angelo Nicolaci, a PhD student at Moffitt Cancer Center. ...evolution. This is a larger process of gradually changing and selecting the best antibodies for a specific...Directed evolution is a method employed to rapidly change a protein towards a specific goal, like improved... end_module_attribute %}{% end_module_block %} Angelo Nicolaci is a PhD candidate in the lab of Jennifer...23(9), 1126–1136. https://doi.org/10.1038/nbt1142 Kang, B. H., Lax, B. M., & Wittrup, K. D. (2022). Yeast... Hilger, D., Rasmussen, S. G. F., Ring, A. M., Manglik, A., & Kruse, A. C. (2018). Yeast surface display...

  2. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    Published
    July 23, 2020, 1:20 p.m.
    ...References Dong D, Guo M, Wang S, Zhu Y, Wang S, Xiong Z, Yang J, Xu Z, Huang Z (2017) Structural basis... 2017 and last updated Jul 23, 2020 by Jennifer Tsang. CRISPR-Cas technology is constantly evolving. Variants...10.1016/j.cell.2017.07.037 Li C, Psatha N, Gil S, Wang H, Papayannopoulou T, Lieber A (2018) HDAd5/35++...//doi.org/10.3390/v10120695 Pawluk A, Amrani N, Zhang Y, Garcia B, Hidalgo-Reyes Y, Lee J, Edraki A, Shah...https://doi.org/10.1016/j.cell.2016.11.017 Shin J, Jiang F, Liu J-J, Bray NL, Rauch BJ, Baik SH, Nogales ...

  3. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    ...biochemical activity Article contributed by Angela Holmes Jin Zhang’s lab is developing new super-resolution...conformational change that placesTagRFP-T and Dronpa in close proximity. By monitoring changes in TagRFP-T... to blue light, CRY2 undergoes a conformational change that permits it to bind to the CIB1 (cryptochrome-interacting...optogenetic system (OptoSTIM1), caused two-fold faster changes in intracellular Ca2+ levels than the original ...Synthetic decaffeination operon Article contributed by Angela Kaczmarczyk  Listen to the synthetic decaffeination...techniques for live-cell imaging of the dynamic changes in biochemical activity within the cell. Their ...

  4. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    Published
    Oct. 11, 2018, 12:30 p.m.
    ...cutting results in 5’ four base overhangs complementary to the overhangs of the assembled inserts. After...in an orientation that results in 5’ four base overhangs after cutting. The destination vector receives... backgrounds – so much so that there was little range for detectable improvements in the methodology. ...Gate assembly, ligation of mismatched pairs of overhangs can lead to incorrect assemblies, so care must... ligation by this enzyme for all 3- and 4-base overhang sequences under standard reaction conditions. ...identification of mismatch-prone pairings. Using these 4 bp overhang observations, accurate “high-fidelity” junction...can be applied to derive similar high-fidelity overhangs for any Golden Gate assembly design. Number...

  5. Addgene's Website Update

    Type
    Blog Post
    Published
    April 8, 2024, 2:15 p.m.
    ... The biggest change is to our most frequently used feature: the search bar. We’ve changed the colors and...Starting today, you may notice some changes to our website! Part of sharing science and making access... development, and content teams have made some changes to the look and feel of addgene.org. Some you’ll...away, while others may be less obvious — but every change is meant to make your experience on our website... on “Deposit Materials” and the search bar will change to an easy-to-use depositing guide, with links ...antibody of the day While those are the biggest changes, there are plenty of less obvious ones to discover...

  6. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ...PMC2629495. Zingg, B., Chou, X., Zhang, Z., Mesik, L., Liang, F., Tao, H.W., and Zhang, L.I. (2017). AAV-mediated...24694006. PubMed Central PMCID: PMC4137353. Chan, K.Y., Jang, M.J., Yoo, B.B., Greenbaum, A., Ravi, N., Wu, W...PubMed PMID: 18223548. Howard, D.B., Powers, K., Wang, Y., and Harvey, B.K. (2008). Tropism and toxicity....A., Ng, L., Winslow, B., Cain, N., Mihalas, S., Wang, Q., Lau, C., Kuan, L., Henry, A.M., Mortrud, M....Wohnoutka, P., Gerfen, C.R., Koch, C., Bernard, A., Dang, C., Jones, A.R., and Zeng, H. (2014). A mesoscale...Hisey, E., Tanaka, M., Kearney, M., Chattree, G., Yang, C. F., Shah, N.M., and Mooney, R. (2017). Identification... 195–206. PubMed PMID: 17343566. Tervo, D.G.R., Huang, B.-Y., Viswanathan, S., Gaj, T., Lavzin, M., Ritola...

  7. Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2:  Reprogramming and Transdifferentiation

    Type
    Blog Post
    Published
    Nov. 6, 2018, 1:12 p.m.
    ...to iPSCs and re-establishment after Partial rearrangement from one cell type to another Time and resources...vitro as part of the process. This opens up a wide range of possibilities in clinical situations. In cases...19998274. PubMed Central PMCID: PMC2946078. 16. Zhang, D., et al., Highly efficient differentiation of...21151107. PubMed Central PMCID: PMC3033971. 18. Huang, S.X., et al., Efficient generation of lung and ...21434814. PubMed Central PMCID: PMC3161101. 20. Chang, C.J., et al., Production of embryonic and fetal-like...2010. 5(1): p. e8763. PubMed PMID: 20098701. 31. Tang, Z.H., et al., Genetic Correction of Induced Pluripotent...PMCID: PMC4835250. 32. Ma, L., Y. Liu, and S.C. Zhang, Directed differentiation of dopamine neurons from...

  8. Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research

    Type
    Blog Post
    Published
    July 29, 2025, 1:15 p.m.
    ... Qiao, J., Zang, Y., Gao, Q., Liu, S., Zhang, X., Hu, W., Wang, Y., Han, C., Li, D., & Wang, X. (2025)...the TAM changes based on the species of origin of the TnpB protein, much like how PAM sites change for different...plasmids. Confirm the correct guide insertion by Sanger sequencing using either 92F or M13R as the sequencing...Dash, M., Saha, R., Das, P., Avinash, S., Shih, J., Yang, Y., Nayak, A. K., Baig, M. J., & Molla, K. A. (...

  9. Science Communication for Everyone

    Type
    Blog Post
    Published
    April 16, 2019, 1:05 p.m.
    ...thanks to our guest blogger, Chinmaya Sandangi! Dr. Chinmaya Sadangi is currently a postdoc at the University...post was contributed by guest blogger Chinmaya Sadangi, a postdoc at the University of Toronto. The Addictive...communicating science to non-scientists. Chinmaya Sadangi, a postdoc at the University of Toronto, created...underrepresented minorities to study STEM subjects. Here, Sadangi and his team answer a few questions about The Addictive...while “manipulate” could mean “worked with” or “changed” to scientists. Keep your message to the main three...Germany in 2017. Apart from The Addictive Brain, Dr. Sadangi is an eLife ambassador, SfN community leader, and...

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

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ..., Villiger, L., Zhou, W., Jiang, K., Garushyants, S. K., Roberts, N., Zhang, L., Vakulskas, C. A., Walker...base-to-base conversion in principle, but some changes require multiple steps and enzymes. Prime editing...can encode any of the 12 possible base-to-base changes in a single edit. Prime editing is also more precise...breaks has been widely used to generate precise changes. However, the efficiency of Cas9 cleavage is relatively...al., 2023). Therefore, they decided to evolve a range of specialized prime editors (PEs). PE6a and PE6b...lab! This post was originally written by Jennifer Tsang in October 2019 and updated by Emily P. Bentley ...Chen, W., Suiter, C. C., Lee, C., Chardon, F. M., Yang, W., Leith, A., Daza, R. M., Martin, B., & Shendure...

  11. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    Published
    May 5, 2020, 1:15 p.m.
    ...the zeptomolar range while One-step-SHERLOCK has a sensitivity in the femto-aptomolar range. For more details... milliliter can be detected – in the zeptomolar range Time: ~1 hour Equipment required: 37°C water bath...powered by a 9V battery which uses a Blue LED and an orange filter to detect fluorescence upon cleavage. Highlights...the immobilised target sequences. Upon binding, changes in fluorescence can be visualised. Another approach...nucleotide variations. Also, FnCas9 cleaves in a range of temperatures varying between 10-50°C. Visit Addgene's...Kellner MJ, Koob JG, Gootenberg JS, Abudayyeh OO, Zhang F (2019) SHERLOCK: nucleic acid detection with CRISPR...Laboratory. https://doi.org/10.1101/2020.04.20.052159 Zhang, F., Abudayyeh, O. O. & Gootenberg, J. S. A protocol...

  12. Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior

    Type
    Blog Post
    Published
    March 5, 2019, 1:08 p.m.
    ... map the turnover of Langerhans cells in the dermis and identify that Langerhans cells are replenished...membrane-tethered CFP (cyan). Brainbow 1.1 adds OFP (orange) (Livet et al., 2007). Brainbow uses a Cre/lox ...respectively, cells will fluoresce either red or orange until recombination occurs. Following the development...incorporates the highly photostable fluorophores mOrange2, EGFP, and mKate2 (red) to solve the problematically... to develop a system for tracking the fate of Langerhans cells. In the Ubow mouse, cells expressing YFP...through the proliferation of specific, and not all, Langerhans cells, to create foci of proliferating cells...., Clément, et al. "Multicolor fate mapping of Langerhans cell homeostasis." Journal of Experimental Medicine...

  13. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    Published
    March 1, 2016, 3:30 p.m.
    ..., 343-345. PubMed PMID: 19363495. 2. Wang JW, Wang A, Li K, Wang B, Jin S, Reiser M, Lockey RF. CRISPR... the 5’ ends of the fragment, generating long overhangs which allows the single stranded regions with ...described in a recent publication by the Lockey lab (Wang, et al. 2015). In this case, rather than using a... along with CRISPR was also described recently (Jiang et al. 2015), in which very large fragments of the... it a try! References: 1. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison CA 3rd, Smith HO. Enzymatic...BioTechniques 2015; 58:161-170. PubMed PMID: 25861928. 3. Jiang W, Zhao X, Gabrieli T, Lou C, Ebenstein Y, Zhu TF...

  14. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...Mi-Yoon Chang, Baek-Soo Han, Sanghyeok Ko, Eungi Yang, Kwang Yul Cha, Robert Lanza, and Kwang-Soo Kim.... 2677165. 3. Jia, Fangjun, Kitchener D. Wilson, Ning Sun, Deepak M. Gupta, Mei Huang, Zongjin Li, Nicholas...includes measles and mumps. Sendai transduces a wide range of cell types and replicates in the cytoplasm independent...using EBNA1 based vectors is that the plasmid can hang around for awhile, even after selection is removed...

  15. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    Published
    Feb. 18, 2016, 3:42 p.m.
    ...enzyme, or enzymes that have compatible overhangs or no overhangs after digestion, you will need to use ...with restriction enzymes to generate compatible overhangs that allow them to bind. Ligase is used to make...ended up in the direction you want - usually by Sanger sequencing or further restriction digests. Of course...inserts cut out of it and your new insert from any overhangs cut off of it via their different migration speeds...insert in a single tube allowing the compatible overhangs generated by restriction digestion to anneal to...only one enzyme or used enzymes with compatible overhangs for your ligation, then you will need to verify...appropriately sized insert, you should send it for sanger sequencing using primers that will allow you to...

  16. Viral Vectors 101: Biosensors

    Type
    Blog Post
    Published
    July 6, 2023, 1 p.m.
    ...fluorescent output, which will turn on, change color, or change intensity of fluorescence in the presence...receptor, but readouts could involve fluorescence, pH change, heat, etc. The last component of the biosensor...engineered to detect a wide array of neurotransmitters ranging from dopamine to glutamate, all the way to acetylcholine...fluorescence-based and have been optimized for a wide-range of uses to monitor the fundamental cellular building...actinin will produce different signals based on changes to the structure of actinin in low vs. high tension...real-time cellular reactions to stimuli and as well as changes in cellular dynamics during so-called housekeeping...

  17. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    Published
    Sept. 15, 2020, 1:15 p.m.
    ...Figure 2: The steps of colony PCR.   Sanger sequencing Sanger sequencing determines the precise order... you'll want to design a custom primer. Because Sanger sequencing can usually sequence just 1 kb of DNA...post for tips for analyzing and troubleshooting Sanger sequencing results. At Addgene, we now use next-generation...that NGS verification is more time-intensive than Sanger sequencing or the other techniques mentioned above...Learn more about Sanger sequencing Read the Molecular Biology Reference ... Jennifer Tsang...

  18. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    Published
    Feb. 24, 2015, 7:50 p.m.
    ...CRISPR technology has changed how scientists edit and control genes, but according to the Broad Institute's...December, Konermann and her colleagues in Feng Zhang’s lab describe just such a system, which they call...perfect complement for one conducted earlier by the Zhang lab for GeCKO, a lentiviral CRISPR library enabling...enabling genome-scale, knockout screening). As the Zhang lab's Nature paper reports, “Here we have shown ...in the genome for gene control over a 1,000-fold range. Weissman explained that there are two levels of... Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O. Cell. 2014 Feb 27;156(5):935-49. doi... Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F. Nature. 2015 Jan 29;517(7536):583-8. doi: 10.1038...

  19. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    Published
    Dec. 17, 2020, 1:30 p.m.
    ...transformed the palette of experiments possible in a wide range of organisms and cell lines. In C. elegans, one ...at 25 ºC. This method resulted in efficiencies ranging from 11-100% of F1 animals carrying precise knock-ins...This stringent selection allowed me to optimize a range of editing parameters: oligo repair templates with...injected animal.  PAM sequence considerations For changes 5’ to the PAM, sequences corresponding to the same...locus (CRISPR-ChAP-MS) could further transform the range of experiments imaginable in C. elegans. Equally... and systems, allowing rapid progress in a wide range of fields.   Thank you to our guest blogger! Jordan...//doi.org/10.1534/genetics.118.301532 Ebbing A, Shang P, Geijsen N, Korswagen HC (2017) Extending the ...

  20. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    Published
    Oct. 8, 2020, 1:15 p.m.
    ...through filtration methods because of its ability to change shape and the absence of a rigid cell wall. Here.... For testing, the medium should not have been changed for at least two or three days before sampling ...commonly infecting mycoplasmas are used. A broad range of primers encompasses almost all the infecting ...chemical reactions and this induces a noticeable color change. This method has high sensitivity but low specificity...are working without gloves or masks. Make sure to change the gloves and masks regularly instead of keeping...water provided to maintain the humidity should be changed regularly as mycoplasma, bacteria, and fungus can...contamination, easy to use, and effective for wide range of mycoplasma. These methods cannot guarantee complete...
Showing: 81 - 100 of 704 results