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We narrowed to 168 results for: plasmid for e coli

Showing: 31 - 60 of 168 results
  1. Plasmids 101: The Wide World of Natural Plasmids

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    ...toolbox. Natural plasmids, also called environmental plasmids, are all around us! A plasmid is a small piece...Old Solution but a New Problem Plasmids 101: What is a Plasmid? Plasmids 101: Transformation, Transduction...Gateway cloning relies on genes found on an E. coli fertility plasmid that also encodes a toxin, CcdB, and the...toxin, CcdA. Knowing which strains of E. coli contain this F plasmid is critical for successful Gateway ...tip! Historically, colicin-producing plasmids have been referred to as “Col plasmids” or “Col factors.”...F. (2023). Plasmid classifications. Plasmid, 126, 102684. https://doi.org/10.1016/j.plasmid.2023.102684...., 2001) Beneficial traits of natural plasmids Natural plasmids often convey a variety of functions that...
  2. CRISPR 101: Multiplex Expression of gRNAs

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    ...on a separate plasmid. Multiplexing in E. coli Koffas Lab CRISPathBrick Multiplex Plasmid: This system ...please scroll down for plasmids suitable for other model systems, including Ecoli, plants, Drosophila,...or SLIC cloning methods. A BsaI-based E. coli multiplexing plasmid is available from the Koffas lab. Scaling..., C, AB, AC, BC, and ABC). Nielsen Lab E. coli CRMAGE Plasmids: The CRMAGE system is a fast, multiplexable...specific point mutations in E. coli. gRNAs are delivered on a different plasmid and allows simultaneous multiplex...Repression in E. coli.” ACS Synthetic Biology 4(9) (2015):987-1000. PubMed PMID: 25822415. Find plasmids from ...multiple gRNAs on the same plasmid, you’ll make sure that each cell that gets the plasmid contains all of the...
  3. R Bodies: Membrane-Rupturing Microscopic Tools

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    ...About Other Hot Plasmid Technologies Read Other Synthetic Biology Posts  Suggest Plasmid Tools for the ...Resources on Addgene.org Read our Hot Plasmids Articles Find Plasmids for Bacterial Expression ... response to changing pH, and they can rupture E. coli spheroplasts to release encapsulated proteins. ...in a single operon and were first expressed in E. coli in the 1980s  (Kanabrocki et al 1986). Much later...showed that these fluorescent R bodies can rupture E. coli spheroplasts (cells which have had their cell walls...Absence of toxicity of R-body producing recombinant E. coli on paramecia. European Journal of Protistology ...produced R-bodies that could extend at different pHs. Plasmids expressing expressing both wild type and mutant...
  4. Magnetic Control of Proteins: More than a Dream

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    ...resources on addgene.org Find Optogenetics Plasmids Find Biosensor Plasmids Visit the Fluorescent Protein Guide...delight,” the very first protein they tested — E. coli expressing good ol’ EGFP — showed a slight but ...1: Fluorescence intensity of EGFP-FlavinTag in E. coli changes in response to a ~25 mT magnet. Gray regions...magnetoresponse was clearly visible whether expressed in E. coli, in mammalian cells, or in purified form, and didn...license.   Video 1: Fluorescence movie of E. coli expressing MagLOV while a magnet is waved under...MagLOV plasmid here!   References and resources References Duret, G., Polali, S., Anderson, E. D., Bell...Frank Hayward et al., 2024). They deposited their plasmid with Addgene, hoping to inspire others to explore...
  5. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

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    ... knockout E. coli strain to assay for functional decaffeination in cells. The ΔguaB E. coli are missing...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...write about a recent plasmid deposit please sign up here.     Listen to the Hot Plasmids episode! Expanding.... Various soCoChR plasmids described in this work including the soCoChR-GFP plasmid as well as several...putida CBB5 and has been tailored for expression in E. coli. This operon is under the transcriptional control...Janthinobacterium sp. Moreover, a caffeine “addicted” E. coli strain expressing this operon can serve as a living...
  6. Plasmids 101: Gateway Cloning

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    ...to find Gateway plasmids for your organisms of interest: Find Gateway cloning plasmids here! References...performed, the next step is to transform competent E. coli cells and select the positive clones. The entry... expression clone. You will also need to use a E. coli strain sensitive to CcdB (e.g. DH5α, TOP10, Mach1... protein inhibits the growth of CcdB sensitive E. coli strains, most colonies should contain the desired...available for all kind of organisms from mammals to E. coli. Glossary of Gateway cloning vectors Vector ...antibiotic resistance markers (indicated here by plasmid color), allowing you to easily select for the expression...desired, recombined construct. Read our recent Plasmids 101 Post on CcdB for more information. How to clone...
  7. Simplify Cloning with in vivo Assembly

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    ...into Ecoli and plasmid assembly (re-circularization) occurs in vivo. We use XL-10 Gold E. coli, but ... assembly of a circular plasmid in vitro, before transforming it into E. coli for propagation. However...blog posts on plasmid cloning Resources on Addgene.org Read our protocols for plasmid cloning Find our...successful recombination reported in all laboratory E. coli strains tested so far. The use of this pathway ... from the MRC Laboratory of Molecular Biology. Plasmid cloning is an essential part of any molecular biology...However, while not widely known, plasmid assembly can be achieved in vivo using a bacterial recombination pathway...can be encoded in the primer sequences. Multiple plasmids can be included in a single PCR, for example for...
  8. Plasmids 101: A Brief History of Plasmids and an Improved eBook!

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    ...tetracycline resistant plasmid, pSC101, and a newly developed kanamycin resistant plasmid, pSC102, with EcoRI...engineering, cloning, and plasmid tags, addtional content added to the "Plasmids that Glow" and "Viral Expression...Learn more about the history of plasmids and cloning: CSHL Meeting: Plasmids: History & Biology The Joshua...Lederberg in 1952 was noted to integrate into the E. coli chromosome in some cases. This terminology held...EcoRI and selected for E. coli transformants that were resistant to both. When this proved successful, ...factors played within an organism.  So how did plasmids get their name? In 1952, Joshua Lederberg set...inheritance factors. He proposed the catch-all term “plasmid” derived as a hybrid of "cytoplasm" and "id" (Latin...
  9. Plasmids 101: Protein Expression

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    ...glycosylated proteins that cannot be produced in  E. coli or yeast cells. The only problem with baculovirus...bacterial host cell is almost always the answer. E. coli is definitely one of the most popular hosts for...expression may be required if your protein is toxic to E. coli. Of all the vectors, pET, under the control of ...modifications Learn more molecular biology in our Plasmids 101 eBook! Researchers use various techniques ...for your protein of interest is inserted into a plasmid expression vector that is then transformed into...being able to help the scientific community share plasmids.   Additional Resources Thermofisher Protein ...Systems Recombinant protein expression in Escherichia coli: advances and challenges Production of recombinant...
  10. Plasmids 101: Blue-white Screening

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    ...Common Lab E. coli Strains Learn to Use Yeast Two Hybrid Systems Browse All of Our Plasmids 101 Posts ...This post is part of our ongoing Plasmids 101 series. Plasmids 101 will provide you with an overview ...of the Plasmids 101 posts here. Now that we have covered antibiotic selection here at Plasmids 101, we...α-complementation plasmid. This way you can quickly and easily identify colonies that not only have your plasmid (antibiotic...rescued by genes supplied on the transformed plasmid. Such plasmids may contain genes that allow for the use...remain the lovely whitish-cream color of standard E. coli. On the other hand, intact β-galactosidase produces...restriction digest. Make sure you use a proper E. coli strain (i.e. contains the lacZΔM15 mutation): XL1...
  11. Harnessing Bacterial Toxins for Allelic Exchange

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    ...Improvement of pCVD442, a suicide plasmid for gene allele exchange in bacteria." Plasmid 51.3 (2004): 246-255. PubMed...one of eight bacterial toxin genes isolated from E. coli or Psuedomonas aeruginosa. These toxins were a ... Gram-negative Enterobacteriaceae including in E. coli O157:H7, Shigella flexneri, Enterobacter cloacae...exogenous enzymes, require multiple transformation and plasmid curing steps, and can generate off-target mutations...a mutated allele introduced from a conjugative plasmid replaces the chromosomal copy. This is a versatile... These two steps are (Figure 1): The initial plasmid integration step (the so-called “single crossover... an allelic exchange experiment? Find the pTOX plasmids here! Continued development of new allelic exchange...
  12. Plasmids 101: Yeast Vectors

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    ...types of yeast plasmids are defined below: Yeast Integrating plasmids (YIp): These plasmids lack an ORI...our first few Plasmids 101 posts, we focused mainly on the elements required for plasmid maintenance within...not only the number of plasmids found in each cell, but also whether the plasmid gets integrated into the...integration. Yeast Episomal plasmids (YEp): These are most similar to bacterial plasmids and are considered “... within an E. coli cell, but vectors can be widely utilized across many different cell types and each ...necessary components to allow shuttling between E. coli and yeast cells. To be useful in the lab, the vectors... Can this be used for auxotrophic selection in E. coli? Additional considerations HIS3 L-hisitidine ...
  13. Addgene's Top 10 Blog Posts of 2015

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    ...Addgenie A. Max Juchheim in this post. 3. Plasmids 101: E. coli Strains for Protein Production Addgenie ...CRISPR mediated genome engineering. 10. Plasmids 101: Control Plasmids Need help thinking about the types ...Scientists eBook Download the Plasmids 101 3rd Ed. eBook Read the Most Recent Plasmids 101, CRISPR 101, and Career...explains why you might want to use a specialized E. coli strain for protein production and gives a quick... two eBooks (Management for Scientists and the Plasmids 101 2nd ed, the 3rd edition has now been released...top of the many developments in science and the plasmids, technologies, and researchers behind them. As...most popular posts continue to be those from our Plasmids 101 and CRISPR 101 series. These posts aim to ...
  14. Plasmids 101: 5 factors to help you choose the right cloning method

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    ... transform the PCR products into E. coli. Ligases within the E. coli then fuse together the PCR products... Read other Plasmids 101 blog posts New to plasmids? Read our article on What is a Plasmid? Download the... the Plasmids 101 eBook Resources on Addgene.org0 Check out protocols for plasmid cloning Find vector...inexpensive DpnI enzyme. Otherwise you let your E. coli do the work for you. Traditional restriction enzyme...
  15. What's the Best Way to Elute and Store Your Plasmid DNA?

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    ...gene endA. Nowadays, the most common E. coli strains used for plasmid amplification are endA negative, meaning...fragmenting plasmid DNA, PCR product, or chromosome. One of the main causes of degradation during plasmid DNA ... a kit Read our blog posts about plasmid cloning Browse our Plasmids 101 blog posts Resources on Addgene.org...Biology Reference Find protocols on plasmid cloning Find plasmids for your research ...DNA extraction and purification from E. coli is the presence of the protein endonuclease I encoded by the...degradation and which experiments you can use your plasmid.  DNA elution choices: TE, Tris buffer, or water...polymerases. In the situation where you need to use plasmid in TE for any enzymatic reaction, an effective ...
  16. Plasmids 101: Shuttle Vectors

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    ... Resistance Gene Plasmids 101: Yeast Vectors Plasmids 101: Mammalian Vectors Plasmids 101: The Promoter...other plasmids? If we all use the same molecular code, can I transfect any old bacterial plasmid into ...blog Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection Plasmids 101:...vectors are usually designed to shuttle between E. coli and Agrobacterium tumefaciens, a plant-infectious...species, for example, in both S. cerevisiae and E. coli. Auxotrophic selection is especially common in ...shuttling between. For example, if you are using E. coli simply for easy cloning and quick DNA production..., where DNA replication begins, so a bacterial plasmid needs a compatible ori that will be recognized ...
  17. Viral Vectors 101: Preparing Pooled Libraries

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    ...library is transformed into E. coli for amplification. Transformed E. coli are allowed to grow on plates...library. This is similar to amplifying a plasmid: transform E. coli with the pooled library DNA, allow the...single plasmid transformation, though, a pooled library has hundreds to millions of distinct plasmids, all...anyone who has worked with individual plasmids knows, some plasmids amplify well and others don’t. The size...pooled library, which contains a collection of plasmids in a single tube, or a pre-made viral prep library... prepare the libraries for you.  Starting from plasmids requires amplification, verification, and lentiviral...library and appropriate packaging and envelope plasmids into HEK cells allows production of lentivirus...
  18. Tips from the Repository Trenches: Using Barcodes to Track Samples

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    ...stored, it is put into E. coli strains that have been treated to easily take in plasmids (transformation). ... of each plasmid so that we never run out. There are a number of different strains of E. coli that can...bacteria with the plasmid can grow. As the bacteria divide, they replicate and retain the plasmid. This allows... Inside Addgene Posts! Learn all about Plasmids in our Plasmids 101 series! ...can be used for transformation. The preferred E. coli strains for storage and distribution at Addgene are...Every plasmid sample enters Addgene the same way. A package is delivered by a mail courier and then the...about Developing Lab Management Software    The plasmid's journey begins First, each package is opened and...
  19. GCE4All: Making Genetic Code Expansion Accessible

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    ...often incorporated into the cells via plasmid. Third, a second plasmid will be needed to introduce the gene... the late nineties, this was developed into an E. coli system, with the noncanonical amino acid in the...expression of phosphoserine-including proteins in E. coli (Zhu et al., 2022). It uses the TAG codon for the...and synthetase introduced to the bacteria using plasmids. Rapid expansion occurred, with hundreds of new...the cell’s genome, instead of incorporation via plasmid or media. Theoretically, if you put all these together...Blue Flame award to identify highly requested GCE plasmids. But they’d also love scientist interested in,...and resources, and then checking out all of the plasmids and bacterial strains (available at Addgene for...
  20. Antibodies 101: Affinity Tags

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    ...Protein as a Solubility Enhancer. In P. E. Vaillancourt, E. coli Gene Expression Protocols (Vol. 205, pp...chromatography. Purification of His fusion proteins from E. coli is generally more straightforward and of a higher...more!   Resources and References   References Coligan, J. E., Dunn, B. M., Speicher, D. W., & Wingfield,...Antibody protocols  Resources on the Addgene blog Plasmids 101: Protein tags Antibodies 101: Epitope tags...
  21. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

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    ...Addgene.org Find plasmids from the Bethune Lab Find Plasmids from the Roux Lab Find Plasmids for FRET ...Split-BioID is BirA*, a promiscuous version of BirA, an E. coli biotin ligase. In BioID, BirA* is fused to a bait... protein complementation assay where BirA*, an E. coli biotin ligase is split into two non-functional ...-BioID Plasmids Here! Munter, S.D., Görnemann, J., Derua, R., Lesage, B., Qian, J., Heroes, E., Waelkens...BirA* fusion proteins are expressed in cells via plasmids or by the creation of stable cell lines and these...Interested in using Split-BioID with your research? Plasmids are available from Addgene! References Schopp...site than in Schopp et al. Find these Split-BioID Plasmids Here!   Burke, B., Kim, D.I., Roux, K.J., & Raida...
  22. FastCloning: A Newer, Simpler Cloning Technique

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    ... Blog Plasmid Cloning by PCR Perform Site Directed Mutagenesis by PCR Catch up on Your Plasmid Background...transformed directly into chemically competent E. coli and transformants selected with the appropriate... have been isolated from methylation competent E. coli for this procedure to work properly. Directly transform...appropriately sized product, purify and sequence the plasmid to confirm no mutations were introduced during ...Background with our Plasmids 101 Posts Resources on Addgene.org Find Empty Vectors for Your Research Find...Browse Our Special Collections for Plasmids from Your Field ... plasmid cloning...
  23. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

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    ... resources on the Addgene blog Plasmid 101 Protein Expression Plasmid 101 Biotinylation Other resources...Burke Lab (Roux et al., 2012). BioID utilizes the E. coli biotin ligase, BirA*, with a site mutation (R11G...-binding domain (233 amino acids) found in the E. coli version. It also contains a single mutation (R40G...to noise ratio 30-fold compared to the standard E. coli BirA* mutant. RapID consists of this mutant BirA...vivo studies (Che and Khavari, 2017). Find APEX plasmids at Addgene! Since the development of BioID and... specific locations in the cell.   Find BioID2 plasmids here APEX2: A more sensitive ascorbate peroxidase... They named this mutant APEX2.  Find the APEX2 plasmids! TurboID and minturbo: Rapid and non-toxic proximity...
  24. Plasmids 101: Gram Negative and Positive Bacteria

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    ...blog Plasmids 101: What is a plasmidPlasmids 101: Blue-white screeningPlasmids 101: Common lab E. Coli... gram negative bacteria include salmonella and E. coli. Because of their thicker peptidoglycan layer, ...useful in a laboratory setting. When designing a plasmid to be inserted into a specific bacterium, it can...it is being used in, and how components of the plasmid should change based on bacteria type. The gram ...
  25. Synthesized by Ginkgo Bioworks, Shared by Addgene: SARS-CoV-2 Plasmids for Many Expression Systems

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    ...first deposited set of plasmids containing SARS-CoV-2 protein coding genes in E. coli and S. cerevisiae expression...What SARS-CoV-2 plasmids from Ginkgo Bioworks are available for request? Currently, plasmids containing all...available. Plasmids are continually added as they complete QC and you can find all available plasmids listed...to fresh data! These same plasmids are then added into our normal full plasmid NGS queue in order to bring...pipeline. For the first ~50 plasmids we perform our standard full plasmid sequencing by NGS. Addgene’s...on Addgene.org Find all plasmids from Ginkgo Bioworks  Find COVID-19 plasmids for your research Browse... all SARS-CoV-2 protein coding genes in E. coli and S. cerevisiae expression vectors are available at ...
  26. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

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    ...recommended for plasmids up to 25 kb, most cloners should be fine, especially using plasmids in the 10-12...Other Plasmid Cloning Techniques like SLIC and Golden Gate Cloning Learn How to Verify Your Plasmid Read...Reference Pages Choose Your Perfect Plasmid Backbone Find Plasmids for Your Research ...nonmethylated PCR products are then transformed into E. coli to undergo nick repair. This method can work well...relying on in vitro assembly, they could make the E. coli do the work by using the principles of recombineering...made genomic SDM relatively straightforward, but plasmid-based SDM has lagged behind. While commercial kits... REPLACR (Recombineering of Ends of Linearized Plasmids After PCR). Current site directed mutagenesis...
  27. Special Delivery: Fluorophore Targeting for FRET Studies

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    ... acids E. coli and Mammalian Cells pEVOL-pAzF synthetase for p-azido-l-phenylalanine E. coli pEVOL-pBpF...cell types (E. coli and Xenopus oocytes, mainly). Nevertheless, tRNA suppression plasmids are available...Resources on Addgene.org Find Plasmids for FRET Browse Empty Plasmids to Fluorescent Protein Fusions ...-pBpF synthetase for p-benzoyl-l-phenylalanine E. coli pAcBac1.tR4-MbPyl pyrrolysyl-tRNA synthetase ...the potential rewards could be substantial: Plasmid tRNA Synthetase Expression Host pMAH-POLY Polyspecific...Fig. 1) which can be carried out using Addgene plasmids. His-tag labeling reagents You may already have...
  28. Plasmids 101: Dimers and Multimers

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    ...recorded plasmid multimerization in the naturally occurring E. coli plasmid ColE1 (Goebel & Helinski, 1968...Addgene blog:  Plasmids 101: What is a plasmid? Plasmids 101: Origin of ReplicationPlasmids 101: PCR...process known as plasmid multimerization. Plasmid Multimerization Early studies on plasmid topology recorded...ligate the plasmid, and re-transform the ligated plasmid to recover the monomeric form of the plasmid. Have...multicopy plasmids harboring large inserts are often dimers in Escherichia coli K-12. Plasmid. 1989 Mar... formation of plasmid multimers (Bedbrook & Ausubel, 1976; Fishel et al., 1981). E. coli strains such ... monomer of Plasmid 12259: pMD2.G (depicted on the left) is ~5.8 kb in length. The plasmid can exist on...
  29. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

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    ...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...promoter.  They found that the system is active in E. coli and closely related species, is amenable to use...be adaptable to distantly related species from E. coli. This tool should serve as both a powerful tool...need. If you'd ever like to write about a recent plasmid deposit please sign up here.        A retroviral...treatment of cardiovascular disease. Find the LipoGlo plasmids at Addgene! Lawler et al, QJM: An International...promoter systems. Find these CRISPR activation plasmids at Addgene. Liu et al., Nature Communications,...
  30. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

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    ...this case, plasmid copy number is the input and gene expression is the output. The input, plasmid copy number...to decouple gene expression and plasmid number. In a suite of plasmids modified from pSC101 with copy ...of all types of plasmids, the stabilized promoters were also tested in different plasmid backbones that...stationary phase, the number of plasmids per cell increases 4-5 fold (for pUC plasmids) or ~2 fold (for p15a, ...RepA is a plasmid replication initiator protein, they could alter copy number of the plasmid by altering...genome or a plasmid. Thus, a stabilized promoter can allow a system to be moved from a plasmid to the genome...special collections page Download the Plasmid 101 eBook See all plasmids deposited by the Voigt lab ...
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