We narrowed to 888 results for: URE

... Addgene scientists inspect each sample and make sure they have been barcoded with a unique ID number ...number (1D barcode as shown below). This number ensures that every sample can be tracked easily at each step...involves good aseptic technique and controls to ensure that nothing is contaminated. We use filtered pipet...both of these are scanned into our LIMS system to ensure that they match, helping us to avoid mixing up ...glycerol stocks on Fridays. From each overnight culture, we prepare three sets of 2D barcoded tubes. Two...receiving, transforming, and storage process, we ensure that the sample tubes that we use match those of... to help track samples. They will help your lab ensure that sample ID’s are unique, consistently labeled...

...systemic biases which, in the absence of support and structure, prevent science from attracting and keeping diverse... diverse talent in science related careers. Failure to strengthen the talent pipeline sets the world up...to be successful after their training.   Figure 1: Professional Development Hub Workshop Attendees... with such a group dedicated to providing the structure and know-how to integrate career development in...areas of the scientific training and employment infrastructure. A Stakeholder Workshop at Janelia Research...2019 solidified the project and the findings were captured in this just released report “Enhancing Dissemination...attracting and launching science talent we must ensure that career skills training is a required and integral...

...included a glossary at the end of this post! Figure 1: Base transitions are edits between the purine...proof of concept for further development. Figure 2: Mechanisms of deaminase-based adenine transversion...for base editor development going forward. Figure 3: Mechanism of gGBE. Created with BioRender.com...•G base editing in mammalian cells and embryos. Nature Biotechnology, 42(4), 638–650. https://doi.org/...editor by engineered human uracil DNA glycosylase. Nature Communications, 15(1), 4897. https://doi.org/10.1038... editor with an N-methylpurine DNA glycosylase. Nature Biotechnology, 41(8), 1080–1084. https://doi.org...efficient T-to-G and C-to-G editing in mammalian cells. Nature Biotechnology, 42(10), 1538–1547. https://doi.org...

... everyone has their ideas heard.  So how can we assure that all participants feel comfortable sharing ...to encourage dissenting opinions is to foster a culture of inclusivity. Work with and value people who ...your lab/company/team is still working towards a culture of encouraging dissent, the next tip may be particularly...feel they can share opposing opinions as well. I am sure we have all experienced a meeting where one person...may not have the highest position or the longest tenure, but don’t let that stop you from speaking up.  ...them. If you can, set aside time to prepare for future challenges. This doesn’t have to be time-consuming...minutes every month dedicated to thinking about the future will improve your ability to innovate. (Finkelstein...

...mCherry is constitutively expressed. Cells are then cultured and FACS sorted. Image from Mateo Sanchez and ...-7 Optimizing fluorescent tools for labeling structures and compartments in mammalian cells By Angela...fluorescent proteins are often used to label  specific structures, compartments, or specific localization of biomolecules...provide misleading information on where exactly a structure or protein is located within a cell. The Goedhart...encoded fluorescent markers for labeling specific structures within cells. HeLa cells were transfected...and mScarlet-I for labeling a list of specific structures within cells. For example, to improve nuclear...microtubules. Find these fluorescent mammalian cell structure markers plasmids at Addgene! Chertkova, AO, et...

...tools and infrastructure in use at research universities not only today, but also in the future. Reproducibility...choosing an ELN for your research, be sure to consider how its features do and don’t support reproducibility...without access to them, the record of research captured in the ELN is incomplete. Thus, to support reproducibility...for discussion and review. It’s also possible to capture Slack conversations and import them into RSpace...and workflows that went into it. The ability to capture the conversations and associate them with a particular...ability to deposit data that has already been structured in folders or notebooks in the ELN, without having...

...Genome-scale measurement of off-target activity using Cas9 toxicity in high-throughput screens." Nature communications...this holds true in genome-wide screens.   Figure 1: Summary of the advantages of the new approach...effects of guide expression and dsDNA breaks. Figure 2: Non-targeting vs. safe-targeting guides. The...dsDNA breaks caused by CRISPR. See supplemental figure 6 for more details. Safe-targeting guides also ...of off-target cleavage by CRISPR-Cas nucleases." Nature biotechnology 33.2 (2015): 187. Pubmed PMID: 25513782...nuclease specificity using truncated guide RNAs." Nature biotechnology 32.3 (2014): 279. Pubmed PMID: 24463574... Other Resources Watch Dr. Michael Bassik’s lecture on “Multiplexing with CRISPR Screens”    ...

...the success of a drug’s clinical and commercial future, and yet are incredibly difficult to find solutions...The consultants who enjoy their jobs most, get pleasure from taking up such complex challenges, quickly...parts of the job? With fast turnover of projects, pressure to maintain consistently high levels of performance...consulting can sometimes be challenging. The fast paced nature of projects often means working long hours both...using lessons from previous cases to better staff future teams. As with any highly collaborative work place... scientific background, training in reviewing literature, and critical analyses skills were applicable... and equity research firms, biotech start-ups, venture capital firms, and more. Essentially any role in...

...GFPs The availability of the three dimensional structure of GFP, its brighter mutant S65T-GFP, and the ...bfloGFPc1. The plasmids used for determining the structures are now available from Addgene - bfloGFPa1 and...Mitochondrial health, transport and turnover are important features to be monitored to study mitochondrial dysfunction...dysfunction. Easy-to-use tools to monitor these features were lacking until Roberta Gottlieb and Zhen Yan labs...pTRE-Tight-MitoTimer) to show its usefulness in cell culture to report changes in mitochondrial turnover and...against common tumor markers to enhance cancer cell capture. Fierer et al., PNAS 2014 Apr 1;111(13):E1176...molecules and exceed a physiologically relevant exposure in intensity and/or duration. Both of these strategies...

...something doesn’t work, it’s not a failure. It’s an education.   Figure 4: Zenodo page showing the IBEX...reagents that work for our experiments!)     Figure 1: The IBEX Imaging Community front page. That...both their successes (recommended reagents) and failures (not recommended reagents) along with information... directly from the Zenodo dataset.     Figure 2: Screenshot showing reagent entries with associated...discussion with others in the IBEX community.   Figure 3: Screenshot showing recent topics in the IBEX...method for high-content imaging of diverse tissues. Nature Protocols, 17(2), Article 2. https://doi.org/10.1038...

...here and no viral progeny are produced (Figure 1).  Figure 1: Comparison of the life cycle of a lentiviral...recombination or non-homologous end joining (see Figure 2).  Figure 2: Schematic representation of integration...cell but cannot integrate it into the host genome (Figure 1). IDLVs were first discovered by researchers ...continuously expressed (Fu et al., 2013). Using IDLVs ensures transient expression without the risks associated...titering IDLVs, there is no integrated piece to measure! Instead, two commonly employed methods for titering...induced by CRISPR-Cas nucleases in human cells. Nature Biotechnology, 31(9), 822–826. https://doi.org/...

...Spinach2 reveals the dynamic nature of trinucleotide repeat–containing RNA." Nature methods 10.12 (2013): 1219...generated allows for qualitative and quantitative measurement of processes including RNA movement by tagging...many structural properties with DFHBI, but its structure allows for greater contact with its RNA aptamer...in the orange or red channel, depending on the structure of the TO derivative (Dolgosheina et al., 2014...imaging small non-coding RNAs in mammalian cells." Nature communications 9.1 (2018): 656. PubMed PMID: 29440634...transcription using a photostable RNA–fluorophore complex." Nature chemical biology 13.11 (2017): 1187. PubMed PMID...efficient RNA mimics of green fluorescent protein." Nature structural & molecular biology 21.8 (2014): 658...

...insert into your vector of choice as shown in the figure below. To start, you need to have DNA fragments...enzymes is that they can all work at the same temperature, so the entire reaction takes an hour or less...an overlap of 15-40 bp, with a primer melting temperature greater than 48℃. Both Snapgene and NEB have ...fragments have stable single stranded DNA secondary structure, such as a hairpin or a stem loop (as might be...overlapping region between fragments also better ensures correct assembly order of the fragments as compared...’ exonuclease instead of using the exonuclease feature of T4 DNA polymerase, which must be controlled ... these kits may come at a high price, their manufacturers tout great efficiency and low time commitments...

...constant, or Fc, domain. Sure, we all know that the Fc domain provides structure, determines isotypes, and...does! While the Fc domain’s functions, besides structure and isotype determination, don’t generally affect...sorted into three main effector functions.  Figure 1: Fc effector functions in an immune response ...antibody-dependent cellular phagocytosis (ADCP). Figure adapted from van Erp, et al., 2019.    Antibody-dependent...description with the Fc domain as only providing structure and isotype determination, remember that it can...Ig Constant Region Effects on Variable Region Structure and Function. Frontiers in Microbiology, 7. https...

...through the La Vida Scholars summer program. I was unsure whether or not I wanted to major in biochemistry..., who let me know in March that I successfully secured an internship at Addgene! I was eager to accept...took into consideration just how busy I was and ensured that my decision deadline would be extended. I ...reached out to Amy with exciting news. I managed to secure a virtual summer program, teaching me the basics...each intern to complete and present on.    Figure 1: Addgene's summer interns presenting their capstone...crunch and knew that if we were to restart the procedure we would not have enough time. During the following...from the electrophoresis, my manager and I were unsure if there were any successful clones because the...

...model and a source of sdAbs (Figure 2) (Xu, et al., 2021).      Figure 2: Genetically modified mice...found similar antibodies in llamas and alpacas (Figure 1). At the time, researchers were already on a ...nanobodies (Arbabi-Ghahroudi, 2017)!      Figure 1: Comparison of a heavy-chain only antibody and... for in vivo experiments, due to their simple structure    sdAbs in research These advantages have contributed...Single-Domain Antibodies: Historical Perspective and Future Outlook. Front Immunol 8:1589. https://doi.org/...mice and llamas neutralize SARS-CoV-2 variants. Nature 595:278–282. https://doi.org/10.1038/s41586-021...

... a couple of lectures in the undergraduate pharmacology course, like one or two lectures a year. And then.... And make sure to subscribe to the Career Channel of the blog so you don’t miss the future posts in this... the first time. I walk[ed them] through their lecture, giving them advice, [and helped] proctor exams...how to design test questions, and how to design lectures. I really built a course from the ground up.”  ...careers while in grad school And just for good measure, here’s a few other posts about developing transferable...

...techniques with higher efficiency and simpler procedures have been developed. Some of these include, TA...annealed and ligated in vivo. Detailed FastCloning procedure To start the FastCloning process, the insert and...and the resultant products of the reaction in the figure from Chaokun et al., 2011 to the right. After PCR...isolated from methylation competent E. coli for this procedure to work properly. Directly transform chemically...chemically competent cells with the digested mixture. There is no need for further purification before transformation...vector + insert). You should screen 3-4 colonies to ensure that you find the correct product. Once you find...have a better understanding of the step by step procedure of FastCloning, here are some tips for high efficiency...

...carte." Nature Reviews Genetics 12, no. 4 (2011): 231-42. PubMed PMID: 21339765. Methods figures were adapted...purify reprogramming proteins. Table 1: Key Features of Different Methods for Generating iPSCs. One...even after many passages. Ban et al developed a temperature sensitive Sendai virus which is removed by culturing...endogenous pluripotency genes (i.e. Oct4) are other measures of reprogramming. If a retrovirus was used, then...field of induced pluripotency: Recent progress and future challenges." Journal of Cellular Physiology 228...pluripotent mouse stem cells by the piggyBac transposon." Nature Methods 6, no. 5 (2009): 363-69. PubMed PMID: 19337237...minicircle vector for deriving human iPS cells." Nature Methods 7, no. 3 (2010): 197-99. PubMed PMID: 20139967...

...been developed around how they replicate (Figure 2). Figure 2: Baltimore replication classes. Image credit...molecular biology tools, a very useful trait. Virus structure While all viruses have a genome inside a protein... with an envelope are called enveloped viruses (Figure 1). Like bacteria, only a tiny fraction of viruses...viruses are pathogenic (Balloux & van Dorp, 2017). Figure 1: A non-enveloped virus (left) and an enveloped...packaged in separate plasmids and expressed via cells cultured with the vectors. This allows the viral vectors...of Viral Vectors 101 posts about each type, so be sure to check our blog, or our Viral Vectors 101 eBook...