...highly recommend watching this entertaining video created by our friends on the Cambridge 2010 iGEM team ...of homology at their ends, which are typically created by PCR. Then, the fragments are incubated together... homology between neighboring fragments can be created via PCR amplification with primers that contain...typically fairly high, so there is usually no need to screen a large amount of colonies. Excluding the time ...advantages that Gibson assembly does not such as increased efficiency, shorter incubation times, or the ability...

...from Aschauer et al., 2013, Figure 1. Used under Creative Commons Attribution License. Within the nervous...can become methylated over time, resulting in decreased expression (Damdindorj et al., 2014; Nieuwenhuis...from Griffin et al., 2019, Figure 4. Used under Creative Commons Attribution 4.0 International License....vice versa. The total volume needed, however, increases with the size of the region you want to cover ... effects (e.g. recombinase-dependent vectors) increases with the amount of virus you add, so it is strongly...

...generated by SnapGene software.  Thanks to increasing affordability and accessibility, more researchers...by growing seed stocks at 30°C compared to 42°C. Screen multiple colonies to isolate the monomer Sometimes... 1984; Summers et al., 1993). In these cases, screening multiple bacterial colonies using one of the detection...DNA-forming sequences and WRN deficiency independently increase the frequency of base substitution in human cells... self-control and a sense of direction are the secrets of multicopy plasmid stability. Mol Microbiol. ...

...promoters and create sophisticated patterns of UAS-driven gene expression. To further increase the complexity...system into a repressible system, Gossen and Bujard created the tetracycline-controlled transactivator (tTA...bacterial operons. A chimeric transactivator, cTA, was created by fusing CymR to the activation domain VP16. cTA...state, and co-expression of the inducer QF can increase QUAS driven expression by ~3,300-fold in Drosophila...

... is a tumour suppressor. This summer role was incredibly valuable because it was a set duration of only...with the large scale data analysis, as this is increasingly reliant on programming skills which are not ...attempted was more meetings, but this simply seemed to create more bureaucracy. As with any large company (both...detection and diagnosis of different types of pancreatic cancer and highlighting which treatment would... mutations. This is a particular challenge in pancreatic cancer where one drug does not work for all patients...

...understanding of how protein folding works and creating biological tools not found in nature. Dr. Baker... Figure 1: Top7 receives the Nobel Prize. Created with BioRender.com. Top7 gets its name from...approximate, two-dimensional map. The Baker Lab wanted to create a new fold that is not present in nature (at least...simply to be stable and novel, the principles that created it have been applied to numerous other protein ...

...example) that are used to increase or decrease expression of a gene. Screen: A library of gRNAs with a...cell population. The resulting mutant cells are screened for phenotypes of interest. Limitations of CRISPR...

...annual Deck the Lab competition drew upon the creativity of the community, who delivered impressive science-themed... used an impressive variety of lab materials to create their decorations! Despite all the craziness of...first place in the home edition, the MicrobiGals created glow in the dark petri dish ornaments and microbe...December 23, 2020   In second place, Dr. Anne Madden created a microbe advent tree displaying over 25 archaea...

... to screen for positive clones in most cases, and restriction enzyme digest can be used to screen plasmids...strand was degraded. Beta: Beta protects the ssDNA created by Exo and promotes its annealing to a complementary...recombination frequency between 0.1% to 1%, and can be increased to as high as 25-50% by designing oligos that ...recombination than dsDNA substrates, so you should screen more clones when using dsDNA. Have you used the...

...be used in concert with in silico prediction to create a more comprehensive list of off-target editing...of DSBs throughout the genome. However, their decreased sensitivity means that the best option moving ...with a focus on the ways science can be used to create a more just and equitable society. Read more at.... Bell, Charles C., et al. "A high-throughput screening strategy for detecting CRISPR-Cas9 induced mutations...Learn to Use Cas9 Activators Perform Genome-Wide Screens with CRISPR Generate Mouse Models with CRISPR/Cas9...

...my partner Mike and I split the work, shared the credit, and we both won accolades that would launch our..., the Senior Scientist who coordinates all kit creation and quality control efforts with depositors; .... We worked with Andrew, the team’s founder, to create a mission statement in the first few meetings and...becoming part of a team is a really good way of creating and strengthening those social bonds. So never...communication options. Addgene, along with an increasing number of academic labs, uses Slack constantly...

...Each path to science policy is unique and this creates a rich and diverse workforce. Out of all the career...QUESTION. Science policy is the work you do to creatively connect seemingly disparate individuals and ideas...also happen at the state and local level. The big secret about science policy that I didn’t learn until ...bring researchers to Congress to advocate for increased science funding and other issues. Whether you ... field where you have no specific scientific credentials.  But before you even apply to that first science...

...paths. Check out Joanne's Reddit AMA Here are some concrete things you can do to develop those transferable...Manage a project involving multiple scientists, create and stick to timelines Be a mentor in a formal ...If opportunities don't exist in your department, create them and show your leadership skills too. Teach...are always excellent Technical knowledge and creativity Become a “thought leader,” develop a technical...

... drug discovery by big pharma (i.e. HTRF and LanthaScreen) (Degorce et al., 2009), measuring intracellular...are critical in setting up a FRET-based assay or screen (the Addgene blog has an excellent primer on FRET... biomolecules each containing multiple binding crevices that can nonspecifically soak up your fluorophore...donors (Fig. 1). Scores of FP variants have been created, thereby enabling a broad spectrum of donor fluorescence...strategies. FRET experimental design lends itself to creativity and innovation, valuable commodities in any NIH...

...COVID-19 research? We decided to give it a try, by creating a service for COVID-19 researchers. So far, we...to researchers worldwide. “COVID-19 has spread incredibly fast and without any notion of national boundaries...-19. “Open scientific sharing is important for creating research synergies and accelerating the translational... but we believe that by coming together, we can create a significant impact on both clinical care and ...

...including Andrea Radtke, PhD, and Ziv Yaniv, PhD, have created the IBEX Knowledge-Base, a community repository...from the Zenodo dataset.     Figure 2: Screenshot showing reagent entries with associated metadata...others in the IBEX community.   Figure 3: Screenshot showing recent topics in the IBEX discussion ...when designing the IBEX Knowledge-Base, seeking to create an iterative process that is constantly becoming...

... The MORF library is only one of 22 libraries, created from 14 deposits, that became available through...would be remiss not to mention that 2024 is an incredibly special year for Addgene: it marks our 20th anniversary...twenty years while seizing new opportunities to create even greater value for the scientific community...every Addgenie, thank you for twenty years of incredible success, and for believing, as we do, that sharing...

...kit size, it may be tempting to try increasing your input to increase your output. But, exceeding a kit’...their relative qualities and quantities. Image created in BioRender.com. Transformation Grade DNA...that you are left with pure plasmid DNA. Image created in BioRender.com. For the clean-up stage, plasmid...kit.   Do you want eco-friendly options? It’s no secret that labs produce a lot of waste. If you are looking...

... to honor the creativity, passion, and dedication to sharing epitomized by the creators of these plasmids...distribute our plasmids via Addgene. Given the increasing popularity of CRISPR in the plant community, ... to send these small tokens of thanks for the incredible impact our depositors have had on Addgene and...

... at eScience Labs, an educational company that creates hands-on science lab kits and digital curricula...writing is clear, Caitlin’s career plans weren’t concrete when she finished grad school at the University...their research from the bench to the classroom by creating the outline of a college level course based on...

...within content and business.  Joining the Addgene crew!  This pattern was consistent: everyone I spoke ...One of the bigger projects I was tasked with was creating and sustaining a biweekly report of our social...coworkers and that trust then translated into this incredible experience.  Of course, I wasn’t presenting to... research on loads of different best practices, create an email strategy, and make the occasional TikTok...

... then degraded by the cell.4.) The transposase creates a double strand break in the DNA and allows the...transposase gains its name from the fact that it was recreated from a defective transposase found in fish. This...transposases. Improvements have been made since then by increasing the consensus between a greater number of extinct...primary cells. In 2009, Zsuzsanna Izsvak’s lab screened many SB tranposase variants for hyperactivity ...

...reminds us that, although fluorescent proteins are incredibly useful, chemical fluorophores can still be used...Dangerous Viruses Contributed by guest blogger Tessa Cressy from the labs of Dr. Elke Mühlberger and Dr. Rachel...CombiGEM and CRISPR for Combinatorial Genetic Screening CombiGEM is a technology that can be used to quickly...barcode small libraries of CRISPR gRNAs to use for screening experiments. Get the full details on how to make...

...and Benjamin Glick from SnapGene. SnapGene was created to meet a need. While there were software tools...the DNA molecules they were using. SnapGene was created to alleviate these problems through good software...control elements. Development of this tool required creating a database of common features, and devising rules...the beginning or end of the feature. With the increasing popularity of gene synthesis, many researchers...

.... “They took a dividing yeast and did a genetic screen to identify the factors that regulate cell division...Leuven in Belgium. “That’s where I not only got incredibly compelled by wet bench molecular research, but... over the years. “Some of the walls we’ve built create inefficiencies that ultimately make innovation ...science slow,” Vandenberghe says. “The continuous creativity that Addgene takes to find ways to disrupt these...

...who are also students in AP biology classes Creating a podcast about genetics using dog breeds as an...structure, and intent to the messaging - while also creating space for people outside of the primary audience...your team?)  Defining your audience allows you to create common ground and understanding that you can use...Image courtesy of Ron Mader, via Flickr, under the Creative Commons license     ...

... of several SARS-CoV-2 variants coincide with increased vaccination efforts. So, how effective are the...Neutralization levels against spike pseudovirus decreases depending on the variant. The lowest levels of... assay to study SARS-CoV-2 spike variants. “We created a series of spike expressing plasmids which encode...different circulating variants. This allowed us to create pseudoviruses that represent lineages and variants...

...promoter, the gene of interest, and a polyA signal. Created with biorender.com. ITR structure  ITRs are symmetrical... and terminal resolution site (trs) are noted. Created with biorender.com. ITR modifications and types...containing transcription factor binding sites can increase transgene expression and transduction efficiency...terminal repeats reduces rAAV productivity but increases transgene expression. Sci Rep, 7(1), 5432. https...

...RspWYL1 increases both targeted and collateral RNA degradation by RspCas13d. RspWYL1 also increased EsCas13d...Cas13d, both groups used very general bioinformatic screens that looked for a CRISPR repeat array near a putative... WYL-domain-containing accessory protein WYL1 increases RspCas13d and EsCas13d cleavage activity The majority...

...application of an electrical charge to cells, to increase cell membrane permeability and thus transformation...DNA can be accidentally packaged into the newly created phages. Transfer of this DNA to another cell is...Viruses (AAV). Lentiviral and AAV can be used to create both transient cell lines, where a gene of interest...Electroporation on the other hand uses electrical pulses to create transient pores in the cell membrane that genetic...

...product and development teams have worked hard in increasing and improving accessibility on our website.  ... In fact, one of our team goals in 2023 was to increase accessibility to our website through the use of...describe images in online spaces. Alt text is used by screen readers to provide visually impaired visitors with...ultracentrifugation. The viral supernatant migrates through increasingly dense layers until the macromolecule density...

...sequencing by NGS. Addgene’s QC scientists immediately screen these plasmids to confirm they contain the expected...of plates will have select samples prepared and screened by NGS to spot check the samples, the rest will...are requested by scientists, will immediately be screened by our QC scientists to confirm the plasmids contain...By leveraging our core strengths of intensive screening and the use of high throughput platforms, Addgene...

... preserving cell viability. Library delivery Screening libraries, large collections of biological independent...independent variables, are becoming increasingly popular tools with many applications. These libraries ...library cargo delivered are controllable. For many screening applications, it is essential to deliver a single...being explored as the delivery mechanism for increasingly popular nucleic acid-based vaccines. These vectors...

...of Producing Monoclonal Antibodies, 1999). This creates an immortal cell population that will continue ...hybridomas for monoclonal antibody production. Image created with BioRender.com.   The antibody-producing... the genes from the hybridoma into a plasmid to create a recombinant antibody.     Figure 2: Changes...Generation of Hybridomas: Permanent Cell Lines Secreting Monoclonal Antibodies. https://www.ncbi.nlm.nih.gov...

...Welcome to our deep dive series, which aims to increase your understanding and technical proficiency with...temperature drops, single intensity should smoothly increase as the DNA amplicon transitions from single-stranded...finally, drop once more during a final temperature increase. If there is a single amplicon, there should only...c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K. Filutowska; Selma, 2019...can fit in a quick game of Connect Four! Image created in BioRender.   Passive Reference Dyes Whatever...

...it, it just reflects how useful the reagents we created are for the community. More than 100 labs worldwide...need for high-quality service, open data, and increased reproducibility of materials. In March 2022, we... supported by an NIH BRAIN Initiative. We also created a growing library of antibody educational resources...York, Calico Life Sciences. Most of all, we’re incredibly thankful for all the scientists who have joined...

...ions flow into the neuron and causes it to fire. Created with BioRender.com. To make channelrhodopsin-based...information about parental channelrhodopsins to create recombinant channelrhodopsin sequences. The model...fiber-optic cable allows blue light to reach the brain. Created with BioRender.com. In the first experiment...neurons in the VTA, this should propel the mice to increase their nose-poking behavior. Strong nose-poking...

...Voigt: The big promise is orthogonality. You can create an almost limitless number of DNA binding proteins...fundamental. We built a NOR gate because they are used to create any program you can imagine. The Apollo 11 mission...so that genetic designs of that scale could be created. Someone could take on an engineering project including...enough hardware to control hundreds of genes. We’ve created enough regulatory elements, CRISPRs, repressors...

...OpenWorm OpenWorm is an open source project that is creating the “first virtual organism in a computer.” This...and split cGAL systems The Sternberg lab in 2016 created cGAL, a GAL-4 based bipartite expression system...reagent toolkit and plasmid assembly line for the creation of CRISPR targeting vectors that each contains...2016). Using SapTrap, this CRISPR plasmid can be created in a single tube. You can also find additional ...

...observe the effects. Luckily for us, CRISPR is an incredibly versatile system that can be tweaked to target...pairs long. Using a single guide RNA (sgRNA) to create a double-strand break works well because very small...ncRNA locus (Ho et al. 2012). For large-scale screening, sgRNA libraries are useful for targeting thousands...092902 Liu Y, Cao Z, Wang Y, et al. Genome-wide screening for functional long noncoding RNAs in human cells...