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TypeBlog PostPublishedJune 25, 2024, 1:15 p.m.... and an extension sequence containing a primer binding site and a reverse transcription template (RTT)...high-throughput libraries My labmates and I wanted to find a way to predict the efficiencies of pegRNA designs...the RTT overhang, the GC content of the primer binding site, and many more. Once we had our datasets with...experiments Prediction models only become useful once they find their way into the hands of users. For this reason...resources References Akhtar, W., De Jong, J., Pindyurin, A. V., Pagie, L., Meuleman, W., De Ridder, J....B. (2021). Impact of chromatin context on Cas9-induced DNA double-strand break repair pathway balance....
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Plasmids 101: 5 factors to help you choose the right cloning method
TypeBlog PostPublishedAug. 21, 2018, 12:31 p.m....traditional restriction cloning. Sequence and Ligation Independent Cloning (SLIC) could also be a great choice, ...non-recombinant vectors from propagating. The ligation-independent methods like Gibson and SLIC are also quite efficient...you some tips for your next project. But keep in mind that there are many other cloning methods - including...don’t be afraid to explore multiple approaches to find out which one works best for you. Happy cloning ...simplified, purification-free, sequence-and ligation-independent PCR cloning method." BMC biotechnology 11.1 (...Addgene.org0 Check out protocols for plasmid cloning Find vector backbones Read our molecular biology reference... -
Quick Guide to Working with Drosophila Part 1: Getting Started with Flies
TypeBlog PostPublishedJuly 13, 2017, 2:30 p.m.... Is it an evolutionarily conserved gene? Can you find an ortholog in the Drosophila genome? Continue reading...describe how you can make your own mutant flies. Find Drosophila Resources at Addgene The birds and the...to help keep your research moving. Below, you’ll find some great introductory resources. As you dive deeper... deeper into Drosophila research, you’ll find more resources that may be more tailored to your field of.... The Bloomington Drosophila Stock Center at Indiana University is another tremendous resource for the...Drosophila Learn about Cre-Lox Resources on Addgene.org Find Drosophila Plamids Drosophila CRISPR Resources Deposit... -
Fluorescent Proteins 101: Fluorescent Protein Timers
TypeBlog PostPublishedMay 4, 2017, 2:30 p.m....whether as part of protein fusions or expressed independently) originally fluoresce green or blue and, when... (green or blue) within a population of timer indicates the age of the pool: the higher the ratio, the...is dependent on the total expression time but independent of protein concentration. Calibration curves ... red/green fluorescence falls within the linear window for the theoretical timer shown in Figure 1B, you...cell structure, etc). FP timers should therefore find use in studies of animal development where events...Your Fluorescent Protein Resources at Addgene.org Find Fluorescent Protein Timers Browse All Fluorescent... -
Plasmids 101: Optimizing Plasmid Yields
TypeBlog PostPublishedMay 26, 2016, 2:30 p.m....following techniques to increase plasmid yield. Keep in mind that you might need to use multiple techniques for...that a higher volume of lysate passed over the binding matrix could clog the matrix when performing filter...determined for each plasmid/strain combination individually, based on determining the cell density or by...Measuring the optical density of the culture can indicate the number of cells present in the given volume... standard 37℃. This plasmid feature should be indicated in the plasmid information or growth strain instructions...coli Strains Additional Resources on Addgene.org Find Info on How to Quantify Your DNA Learn to Do a Bacterial... -
Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More
TypeBlog PostPublishedJune 12, 2014, 3:30 p.m.... switch to Golden Gate Cloning easier because individual labs will not need to ‘domesticate’ all the basic... they were developed mostly with model plants in mind, Arabidopsis and Nicotiana benthamiana. N. benthamiana...Golden Gate cloning system especially with plants in mind. When it’s ready, I’ll make those available on my... up making 30 constructs. They’ve never done any kind of cloning and they don’t have any prejudice about... active on Twitter. Sometimes it’s a good way to find out what’s happening in the office next to you. ...remarkable number of female scientists whose tweets I find really inspirational. They share experiences, articles... -
Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy
TypeBlog PostPublishedAug. 27, 2020, 1:15 p.m....protein (FP) is split into two fragments that individually lack fluorescence. When complementary FP fragments...resources in less genetically tractable organisms lags behind. BiFC can also be used to screen for small-molecule...carry out because it relies on a single BiFC pair. Find BiFC plasmids for your experiment at Addgene! How... the random association between FP fragments independent of PPIs. Overexpression can be assessed by comparing...fluorescence due to the random association between FPs independent of PPIs. Simply compare the fluorescence intensity... introduction to FRET Resources on Addgene.org Find fluorescent protein plasmids and resources Visit... -
Capturing a Moment with Marc Zimmer’s “The State of Science”
TypeBlog PostPublishedJuly 30, 2020, 1:15 p.m....of genome engineering via CRISPR. Though the individual topics are not surprising, the real value here...Science,” he describes the now infamous tale of Rosalind Franklin and her essential-but-unlauded role in...quieter, more reserved Franklins and Prashers will find it increasingly difficult to compete (for funding... quite a walk down memory lane for me. A brief window in time “The most obvious future consequence of... but that’s unlikely to be foremost in readers’ minds. Six months ago I would have nodded my head knowingly...book left them wanting to learn more. There are individual paragraphs that may have warranted their own ... -
Summer SciComm Series: Modes of Communication
TypeBlog PostPublishedJuly 1, 2022, 1:15 p.m....proficiency. What kind of content are you sharing? Sometimes your subject matter will drive the kinds of communications... I want to put to rest a common misconception. I find personal bias often unintentionally plays a big ...My dad, on the other hand, mastered the art of finding how-to videos on YouTube long before he learned...audience already is. You’ll also want to keep in mind that access and availability to resources (including...embark on a project, do your best to understand what kind of resources - including your time - will be required... -
Plasmids 101: Choosing an Antibiotic Resistance Gene
TypeBlog PostPublishedOct. 10, 2023, 1 p.m....expressing a protein that directly binds to zeocin, preventing it from binding DNA. Zeocin is an all-in-one ...a way to produce more of the plasmid, isolate individual plasmids, or express a gene contained in a plasmid...member of the bleomycin family of antibiotics, which induce cell death by intercalating into DNA and generating...genotoxic when not 100% inhibited, meaning it will induce mutations in the host/plasmid DNA during selection...the Sh ble gene does not fully suppress Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2)... -
Better Dyeing Through Chemistry & Small Molecule Fluorophores
TypeBlog PostPublishedSept. 8, 2016, 2:30 p.m.... determines how long and precise we can track individual molecules. Poor photon budgets are a widespread... synthetic fluorophore-contianing “ligand” that binds to the tag. Like most good ideas in biological imaging...proteins (FAPs) – These modified antibody fragments bind and enhance small molecule fluorogens (Figure 1e... encoded tag is required as the small-molecule binding motif targets endogenous protein (Figure 1f). ...when free in solution, but high fluorescence upon binding to its cognate protein. In some cases this property...Request Plasmids from the Michael Davidson Collection Find Fluorescent Protein Kits ... -
Overcoming the AAV Size Limitation for CRISPR Delivery
TypeBlog PostPublishedSept. 16, 2020, 1 p.m.... above showed successful target modification, indicating that AAV is a good delivery vehicle for Cas9....cleavage activity in mammalian cells. SaCas9 produced indels at a similar efficiency to SpCas9, leading the ...tested two genes in mice. For both genes, they saw indel formation and phenotypic changes as early as 1 week...packaged in AAV. Split AAV approaches While finding smaller Cas9 orthologs works for some applications... in two pieces and package each piece into an individual AAV. When a cell is transduced by both AAVs, ...Resources on Addgene.org Check out our CRISPR Guide Find CRISPR Plasmids ... -
Four Base Editing Reporters to Monitor and Enrich Editing in Real-time
TypeBlog PostPublishedJuly 7, 2020, 1:15 p.m.... To create the eGFP reporters the Harris lab individually mutated three codons in eGFP to eliminate fluorescence...capability of the seven human APOBEC3 proteins, finding that APOBEC3A and APOBEC3bctd are the most efficient...using a base-editor (C>T conversion) they could induce the translation of any detectable protein, not ...contained the cytidine base editor exhibited robust induction of GFP fluorescence confirming the efficacy of... universal, the Dow lab successfully used GO to induce the translation of an array of different reporters...Resources on Addgene.org Browse the CRISPR guide Find CRISPR plasmids for your research ... -
Save Time with Transient Plant Leaf Transformations
TypeBlog PostPublishedJuly 25, 2019, 12:03 p.m....Plant Plasmids and Resources page Find synthetic biology plant plasmids Find CRISPR plant plasmids ...plant. A common method for silencing genes Virus induced gene silencing (VIGS) is a transient technique ...subcellular localization of your protein of interest. Find all plasmids from the Lee-Parsons lab. Test different...PMC6585625. Unver, Turgay, and Hikmet Budak. "Virus-induced gene silencing, a post transcriptional gene silencing...Addgene blog Read all of our plant biology blog posts Find tips for Arabidopsis transformation Read our Plasmids... -
Plasmids 101: Gateway Cloning
TypeBlog PostPublishedJan. 12, 2017, 3:30 p.m....Use the links below to find Gateway plasmids for your organisms of interest: Find Gateway cloning plasmids...carry different antibiotic resistance markers (indicated here by plasmid color), allowing you to easily...pLenti CMV Puro DEST (w118-1) or the doxycycline-inducible pLIX_403. The chosen attR destination vector will...fragments, cDNA or Genomic DNA and is available for all kind of organisms from mammals to E. coli. Glossary of...Quickly Verify Your Clones Resources on Addgene.org Find Plasmid Cloning Protocols Watch Our Diagnostic Restriction... -
5 Reasons to Use Reddit for Science Communication
TypeBlog PostPublishedOct. 20, 2016, 2:30 p.m....forum discussions about a wide variety of topics. Individual forums are hosted on subreddits (r/”topic”) which...yourself and your work out to other scientists who will find it useful, I highly recommend doing an AMA! Before...something we are, not something we do. We have to find a way to keep doing science to be happy. The happy... I said in the AMA, it’s incredibly important to find good mentors and incentivize good mentorship: AMA...get to speak publically about your work (you can find my tips for a perfect science talk here). Finally...the features I need to save, organize and easily find the resources that I come across on the internet... -
Plasmids 101: Broad Host Range Plasmids
TypeBlog PostPublishedApril 11, 2024, 1:15 p.m....et al. 2010), inducible promoters with different degrees of stringency and inducibility (Toh et al. 2023... replication initiation factors and proteins independent of the host. Host range depends on whether the... vary in DnaA-box and iteron structure, host-independent Building broad host range recombinant...2023), and additional cloning sites (Keen 1988). Find broad host range recombinant vectors at Addgene!... (2023). Development of constitutive and IPTG-inducible integron promoter-based expression systems for... -
Data Freedom: The Expansion of Data Sharing in Research Publications
TypeBlog PostPublishedAug. 5, 2014, 6:51 p.m....different or novel tools and assumptions to reveal findings that the originator was oblivious to, no matter...of the experiment would require access to each individual video. To PLOS’s credit, the organization subsequently...recognize that storing data in a sharable form is indeed a significant effort. Our standards of data recording...recording are highly variable and usually left to individual researchers. Yet accurate and comprehensible ...appears to leave the scope of data to be shared to individual authors. Minimally, raw but sufficiently annotated...charges of elitism and for scientists to be hiding behind our jargon and esoterica. Public trust in and funding... -
Streaking for Single Colonies: The Streak Plate Challenge
TypeBlog PostPublishedMarch 2, 2021, 2:15 p.m....from the termite hindgut. Want to level up your or your class’s streak plate skills? Find the chromoprotein...Resources on Addgene.org Find fluorescent protein plasmids at Addgene Find chromoprotein plasmids at ...the students to separate out colonies from each individual strain using the streak plate technique. Time... -
Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus
TypeBlog PostPublishedSept. 1, 2020, 1:15 p.m...., 2008). Shortly after though, three groups independently tried reproducing these results and determined... and part B. Each piece of the gene is then independently packaged into an AAV. When a cell is co-infected...Viral Vector Packaging Capacity in a Transgene-independent Manner. Molecular Therapy 16:124–130 . https:...AAV Transductifon Is Mediated via a DNA-PKcs-independent, Rad51C-dependent Repair Pathway. Molecular Therapy...ddv386 Additional resources on the Addgene blog: Find tips for a first time AAV user Learn about how AAV...on Addgene.org: Check out the Addgene AAV guide Find ready-to-use AAV preps from Addgene's viral service... -
Transferable Skills Guide: Conflict Resolution
TypeBlog PostPublishedJan. 4, 2022, 2:15 p.m....these types of conflicts is important for the individuals involved, and also for the labs, organizations...they were taking care of that”, because that can indicate confusion around... Ownership: Misunderstandings...tools that can come in handy: Empathy: If you often find yourself in confrontational situations with others...the age of five. But it does mean that even if you find someone annoying or disagreeable, you recognize ...Definitely not. But it can help, and if and when you find yourself transferring this skill to another job,...for avoiding, competing, or accommodating, if you find yourself gravitating most frequently towards these... -
Multicolor Animals: Using Fluorescent Proteins to Understand Single Cell Behavior
TypeBlog PostPublishedMarch 5, 2019, 1:08 p.m....copies of the transgene that will recombine independently, cells may acquire one of a variety of colors...extremely versatile technology as recombination can be induced in developing fish via Cre protein microinjection...microinjection or crossing of Zebrabow animals to an inducible Cre line. The Schier group developed Zebrabow as ...powerful tool for understanding cell proliferation. Inducible XFP expression in the Ubow mouse Another murine...CFP, and YFP. After crossing onto a tamoxifen-inducible ubiquitin background, each cell in the mouse that...Köhler, Corinna, et al. "Mouse cutaneous melanoma induced by mutant BRaf arises from expansion and dedifferentiation... -
New Optogenetic Tools for Cytoskeleton and Membrane Control
TypeBlog PostPublishedSept. 28, 2023, 1:15 p.m....single-component systems take advantage of BcLOV4’s light-induced binding of the plasma membrane. In previous publications...light BcLOV4 undergoes a conformational change and binds the plasma membrane under blue light CRY2-CIB1 heterodimerizes...limitations (Figure 1). Figure 1: Opto-katanin induces fast, localized microtubule severing. A) Schematic...substantial PA accumulates in the plasma membranes, indicated by the PA probe colocalizing with SuperPLDx30....CRY2-CIB, and other optogenetic modules? Or want to find more optogenetic tools in our collection? Check ... -
Antibodies 101: Introduction to Gating in Flow Cytometry
TypeBlog PostPublishedApril 4, 2024, 1:15 p.m....complexity inside the cell or on its surface. Bear in mind, the voltage setting of the laser impacts the magnitude...While flow cytometry can be used for virtually all kinds of experiments and research fields, for any experiment...After that, the next gates fully depend on your individual experiment. Practical example: lymphocytes in...cellular debris and other larger cells. Note, we don’t mind the warning message at the top, as our cells of ...strategy, I can, of course, also draw multiple individual gates around the populations of interest (Figure...of a quadrant gate, I am free to use multiple individual gates. Note that in this case, square gates were... -
Site Directed Mutagenesis by PCR
TypeBlog PostPublishedAug. 2, 2016, 2:30 p.m....homology-directed repair (HDR) of a CRISPR/Cas9 induced double-stranded break. This site-directed genome...rendering the resulting construct resistant to Cas9 induced cleavage. Summary of Site Directed Mutagenesis ...is removed by DpnI restriction digestion leaving behind only the mutagenized plasmid. Although it is commonly...reaction. Ideally, your primers should be free of palindromic and repetitive sequences, but if present, a minor...similar approach can be used to identify Cas9/CRISPR-induced genome modifications. Occasionally, multimerization...Addgene.org Browse Our Molecular Biology Protocols Find Empty Vectors for Your Next Cloning Experiment ... -
Using Phosphoserine to Study Protein Phosphorylation
TypeBlog PostPublishedJune 23, 2016, 2:30 p.m....on Addgene.org Find Synthetic Biology Plasmids Find Bacterial Expression Systems Find Plasmids for Protein...optimized for parameters such as induction temperature and length of induction, which will vary between different...gels containing Phos-Tag, a small molecule that binds specifically to phosphate moieties and promotes ... -
Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons
TypeBlog PostPublishedApril 24, 2015, 2:39 p.m....Advances in Cre-lox recombination have included inducible Cre, such as the tamoxifen-responsive CreERT, ...single-cell labeling is necessary to distinguish individual cells. The Brainbow-1.0 construct contains three...mice.) The combinatorial effect of multiple, independent recombination events leads to a rainbow of colors... with the vast array of colors needed to mark individual neurons, the system also suffered from a number...they screened a variety of fluorescent proteins to find those with ideal characteristics (low aggregation...Check out Brainbow plasmids available from Addgene! Find Brainbow Plasmids @Addgene: Brainbow 1.0, 1.1, ... -
Addgene Stands with the Scientific Community
TypeBlog PostPublishedApril 23, 2025, 1:30 p.m....science funding, limitations to facilities and indirect costs, and increased tariffs present an unprecedented... of the cost of creating and verifying them independently — particularly valuable when budgets are constrained...preparations, and antibody production that labs may find increasingly difficult to maintain or access. These...applications, addressing the new funding structures and indirect cost caps more efficiently. Offering free educational... seeking and expanding our collaborations with industry and public and private partners to create additional... -
Inntags: Innovative Protein Epitope Tagging
TypeBlog PostPublishedNov. 10, 2015, 3:30 p.m....similarly to their untagged counterparts. These findings motivated Georgieva et al. to develop new tags...from non-vertebrate systems. Preliminary testing indicated that inntags IT5, IT6, and IT10 expressed well...on these parameters. IT5 and IT6 displayed good binding efficiency in both immunofluorescence and immunoprecipitation...for both applications. To prove that inntags are indeed innocuous, Georgieva et al. created tagged versions...detrimental effects on cell size and growth rate, indicating that they compromised protein-protein interactions... -
CRISPR-Cas14: a family of small DNA-targeting enzymes enabling high-fidelity SNP genotyping
TypeBlog PostPublishedNov. 29, 2018, 1:53 p.m....protection against infectious agents. Several enzymes behind this immunity have already been discovered and ...coli bacteria strain. They found that Cas14 could bind and cleave a targeted sequence of ssDNA. Unlike ...cleavage, the authors showed that Cas14 could indiscriminately cleave ssDNA in trans like Cas13 and Cas12...of dsDNA and RNA in trans. Fluorescent signal indicates the presence of this viral sequence in the assay...our CRISPR cheat sheet Resources on Addgene.org Find the Cas14 plasmids here Checkout our CRISPR guide... -
"What Makes a Good Mentor?" and 6 More FAQs About Science Mentoring
TypeBlog PostPublishedDec. 17, 2013, 2:47 p.m....Advisor/PI who will also be a good mentor? Where can I find a Mentor? What is this I hear about “group” or “...“A good mentor has your goals/needs/situation in mind, rather than (or as well as) their own.” “Someone...mentors/advisors like Yoda? If so, you might want to find additional sources of wisdom. While somewhat lighthearted...on the blog “Tenure, She Wrote” is titled, “What Kind of Mentor Do I Want To Be?” In this excellent essay...the northeast, writes a thoughtful treatise on the kind of advisor/mentor she wants to be for her new lab... -
Using AAV for Neuronal Tracing
TypeBlog PostPublishedAug. 9, 2018, 1:04 p.m....using AAVs Resources on Addgene.org Find ready-to-use AAV Preps Find AAV plasmids Browse all viral vectors... different parts of the brain is mediated via individual neurons. Neurons are composed of a cell body,...axons directly to the prefrontal cortex, but also indirectly via synapses to neurons in other brain regions...and aid in visualization of the morphology of individual neurons, including dendritic and axonal extensions...these neurons fluoresce in different colors. An individual neuron can then be traced throughout the brain...Viswanathan, S., Gaj, T., Lavzin, M., Ritola, K.D., Lindo, S., Michael, S., Kuleshova, E., Ojala, D., Huang... -
Scientific Sharing in the Time of COVID-19: Databases and Resources
TypeBlog PostPublishedMay 12, 2020, 1:15 p.m....have rapidly been developed. We asked the individuals behind some of these new platforms what their motivations...collaborations within the scientific community The team behind Crowdfight COVID-19 strives to match up scientists...ultimately treatments in a timely fashion.” With this in mind, Bourque and Tara Moriarty, an Associate Professor...extremely important part of the future of research. To find other COVID-19 databases and resources, check out... -
Supporting Rare Disease Research with BeHeard 2017
TypeBlog PostPublishedJuly 20, 2017, 2:30 p.m....spinal muscular atrophy, the molecular mechanisms behind the disease are unclear. Dr. Di Schiavi plans on...concrete understanding of the molecular biology behind SMN1 and its interaction partners, Dr. Di Schiavi...suffering from this little-known disease. You can find more information about the disease on the Meothelioma...suffering from rare diseases. These underrepresented individuals deserve a voice. Check back with Addgene often...Countries with Seeding Labs Resources on Addgene.org Find Plasmids for Disease Research ... -
pSiM24: Simplifying Plant Genetic Engineering
TypeBlog PostPublishedSept. 29, 2015, 2:30 p.m....: 4045853. Find other plant resources at Addgene Check out Addgene’s plant vectors! Find the plasmids...Agrobacterium tumifaciens. Agrobacterium carries a “tumor-inducing” or Ti plasmid, which allows it to transfer genetic...during preparation in E. coli. Addgene depositor Indu Maiti has created a new and versatile binary Ti ...References Sahoo, Dipak Kumar, Nrisingha Dey, and Indu Bhushan Maiti. "pSiM24 is a novel versatile gene... -
Cre-ating New Methods for Site-specific Recombination in Drosophila
TypeBlog PostPublishedMay 12, 2015, 1:32 p.m....different constructs and cell types. Second, Cre may induce recombination at pseudo- or cryptic loxP sites ...proliferating cells. A Cre-estrogen receptor ligand binding domain-fusion can prevent this toxicity, but with...offspring carried both the recombinase and GAL4, indicating that the recombinases were inherited normally...10.1073/pnas.1111704108. Epub 2011 Aug 9. PubMed. Find the Plasmids @Addgene: Plasmids for KD, R, B2, ...-665. PubMed. Growth inhibition and DNA damage induced by Cre recombinase in mammalian cells. Loonstra... -
In Living Color: The Skinny on In Vivo Imaging Tools
TypeBlog PostPublishedMarch 27, 2014, 5:14 p.m....maybe a tumor. They wanted to find something better. Transparency window That’s exactly what they reported... Verkhusha explained it to me, the transparency window of mammalian tissues is set by the properties of... all of them fluoresce outside the transparency window of mammalian tissues,” Verkhusha said. “We developed...near-infrared fluorescent proteins within this transparency window, so now, we could see deeper.” Adding to the toolbox... -
Viral Vectors 101: Parts of the AAV Packaging Plasmid
TypeBlog PostPublishedDec. 3, 2020, 2:15 p.m....group needed for DNA replication, Rep78/68 binds the Rep binding element (RBE) in the ITRs, uses its endonuclease...N-terminal domain of Rep78 and Rep68 contains a DNA-binding and endonuclease domains, all four Rep proteins...onto an assembled capsid. Then Rep52/40, which also bind the capsid, “pumps” the genome into the capsid through...plasmid Resources on Addgene.org Learn about AAV Find AAVs for your experiment Browse our AAV protocols... -
Early Career Researcher Toolbox: Free Tools for Making Scientific Graphics
TypeBlog PostPublishedJune 11, 2019, 1:16 p.m....Vectr has layers, which let you lock and hide individual vectors (shapes defined by 2D points connected... open source and available for desktop use for Windows, Mac OS X, and Linux. Inkscape is versatile and...biology, medical specialities, and general items. Individual images as well as collections of images are easy...appropriate credit, provide a link to the license, and indicate if changes were made to the images. This requirement...Toolbox blog post on social media for scientists Find science career blog posts Learn about other lab ... -
Antibodies 101: Affinity Tags
TypeBlog PostPublishedSept. 19, 2023, 1 p.m....most common tags available. Maltose binding protein Maltose binding protein (MBP) is a 45 kDa tag which...system (bacteria, yeast, mammalian, etc.). To individually isolate a protein, there must be a special feature...while epitope tags are used for both direct and indirect visualization. Some of these tags can also be ...0471140864 Fox, J. D., & Waugh, D. S. (2002). Maltose-Binding Protein as a Solubility Enhancer. In P. E. Vaillancourt...