...type; however, their usage has been limited by technical problems. For instance, it’s much more difficult...protein-only transduction method and its potential applications to CRISPR/Cas9 genome editing.   Protein-only...that components in the buffer used for protein purification must be helping the untagged protein enter cells...Diego D'Astolfo. CRISPR/Cas9: a potential application of iTOP Having tested iTOP using Cre-lox recombination...showed up to 70% resistance to diphtheria toxin, indicating a 70% frequency of biallelic mutations. Importantly...Importantly, hESCs retained their stemness, indicating that iTOP may be valuable for CRISPR/Cas9 stem cell...potential for off-target effects. Third, protein purification (once a protocol has been established) may be...

...researcher and science communicator.  Update (7/17/2018): Check out Fatima's recent publication on her HMO work...protein-protein interactions. Beyond these common applications, researchers are finding new ways to apply fluorescent...Zhou and Fatima Enam. There were many excellent applicants but Xin and Fatima stood out for their creativity...utility of these FLiPs in a variety of research applications and particularly notes their ability to be precisely...known to bind to the HMO of interest. Fatima specifically proposes using this sensor to detect HMOs found...endeavors, Fatima hopes to practice her science communication skills throughout her training and wants to ...work in Cell Chemical Biology! Additional Resources on the Addgene Blog Learn More about Photosactivatable...

...Harvard Medical School and the University of Toronto. He is an author of 20+ scientific publications and ...quick turn-around time of Sanger sequencing have historically matched the needs of most molecular biology ...smaller fragments that are then barcoded for identification. This mixture of barcoded fragments is known... a collection of 96 plasmids on a plate) will typically have a wide range of input quantities. This variation...samples. As mentioned above, one of the primary technical benefits of plexWell™ is that normalized libraries... proprietary chemistry). This turns out to be critically important for getting adequate sequencing depth...computational biologist with a passion for solving critical sequencing problems. He received his PhD in Bioinformatics...

...MSDS for a lab chemical of choice. Now, I can’t guarantee every reagent in your chemical closet is in this...lab notebook. Lab Archives is a multipurpose application that allows you to share your data with colleagues...directions including which buffers to use and that indicates if a sequential digest is needed. The enzyme finder...enzyme sequence resource is universally useful. 4. Chemical Safety Data Sheets by ThatsMyStapler Safety always...experiments. Hence, I’m including this MSDS app. The Chemical Safety Data Sheets app has a search function that... if you get quizzed randomly about a hazardous chemical and can’t find your lab’s MSDS book, this is a...AMINO is a very simple app that packs important biological information, amino acid abbreviations, and structures...

...these proteins are FDA-approved or in clinical trials or preclinical studies already. We are following up...expand to other cell types, including more physiologically-relevant systems, to catch all the important... we found in our map have an impact on virus replication. While we and our collaborators are starting ... Nsp3 and Nsp16, and includes an additional catalytically dead version of the viral protease, Nsp5. Our... 2xStrep-tag for easy detection and affinity purification. All but one are in a pLVX vector background...coronavirus, one FedEx package at a time Since the publication of our paper, our group has been inundated with...Eckhardt and Melanie Brewer are on the Science Communications Team at QBI UCSF. They are particularly interested...

...ll be talking about the ins and outs of this classification system, including its history, biology, and...repeatable phenomenon, Gram began using it as a classification system. Eventually, any bacteria that retained...gramstain.html Smith, A. & Hussey, M. Gram Stain Protocols. American Society for Microbiology (2005). https://asm.org... or pink. Purpose Gram staining can be used in medical settings to determine the general type of bacteria...permeable to different classes of antibiotics. The classification system is also useful in a laboratory setting...bacteria type. The gram positive and negative classification system is incredibly helpful in visualizing...stepping into a lab.   Anna Kamens is a science communication summer intern at Addgene and a rising third-...

...Technology and its Application in Genetic Diseases: A Review.” Current pharmaceutical biotechnology. https...2020.  “CRISPR Gene Therapy: Applications, Limitations, and Implications for the Future.” Front. Oncol...several treatments are currently being tested in clinical trials. It is speeding the process of the discovery...the technology in different labs for different applications.” Li’s team noted that CRISPR/Cas9 has been ...cleavage sites are repaired.” Those insights will be critical for scientists’ ability to influence editing efficiency...with size limitations. In addition, non-viral applications are also possible through the use of nanoparticles...

...mentor (if you have one) to communicate your plan as well. Making logistical and funding arrangements Visiting...the host university does not hire or pay them (typically). A visiting scientist is distinct from equipment...the scientist to the new institute where their typical workdays will be carried out until the end of the...equipment! Preparing for your visit   Design and communicate a research plan Your time at the new research... and you are set up for success. If you are communicating with a professor or institute, ask if you will...expect to generate data that will contribute to a publication? A grant? A patent? It is never too early to ...Resources Resources on the Addgene blog Fostering communication in remote collaborations Transferable Skills...

... and 15/17 morphologically aligned. Documentation of the network that initiates biological reprogramming...unique barcodes for each TF help streamline identification. While the Zhang lab controlled their multiplicity...single TF was delivered to each cell, future applications could deliver multiple TFs to each cell, allowing...processes. For help with library transformation and amplification, refer to the Zhang lab’s previously published...published protocol and its relevant modifications on the pooled library webpage. Disease models Good disease...generate, in part because of difficulty with replicating the specific dysregulation in the disease. For...Addgene.org Plasmid Pooled Libraries Pooled library amplification Zhang Lab CRISPR Plasmids The Human TFome Library...

...agreement on the transfer of biological materials in 1995: the UBMTA. Because biological materials are widely ...many different flavors of MTAs, but the Uniform Biological Material Transfer Agreement (UBMTA) is one of...MTA for that plasmid facilitated by Addgene — basically, instead of having to sign individual MTAs for...things being organized, tracked, and clearly communicated. And that’s where MTAs, and similar agreements...agreements allow for centralized tracking when biological reagents are shared — both at what time and with... options for the terms you want to share your biological materials under! While most plasmids at Addgene...portal automates a lot of the process, from notifications and emails to generating the UBMTA link for ...

... Connie Cepko (Harvard Medical School), and Dr. George Church (Harvard Medical School) for taking the ...does one stand out amongst a pool of successful applicants and get chosen to work in his or her lab of choice...environment? Does the group expect each other to be physically present at certain times during the day? We asked... next.” Connie Cepko looks for people who are dedicated and interested in science. “Postdoc hires are ...history in science. Potential graduate students typically go through a rotation, which reveals their working...job satisfaction (especially for scientists and technical people) is linked to engagement and access to ... "chaotic" and offer less micromanaging, they typically offer more flexibility in choosing direction of...

...cost-prohibitive for many labs, and are often impractical for small projects. Most researchers instead ...involves splitting up a sample into thousands of physically isolated partitions (such as droplets or wells...Since there are many thousands of partitions, statistical modelling can be used to determine the actual..., does not require standard curves, and can theoretically detect a single target molecule in a sample....a gene of interest is one of the most common applications of precision genome editing, the remainder of...single molecule resolution of digital PCR allows classification of individual target molecules based on their...single wells of a 96-well plate without prior quantification. Comparatively, mismatch assays require between...

...systems are available at Addgene! Chemically inducible promoters Chemically regulated promoters are among ...sensitive expression systems are typically less leaky than chemically induced promoters; they show near-zero...transcription is turned ON. Types of inducible promoters Chemical agents, temperature, and light are all examples...prokaryotic promoter often used for bacterial protein purification. When arabinose is absent, regulatory protein...decreasing promoter leakiness.   Other examples of chemically induced promoters include positive inducible ... promoters only work in prokaryotic cells and typically only in the same or closely related species from...

...Following up its 2018 iGEM debut - centered around a prototypical methane biosensor - IISER Bhopal is back in ...psychrophilic enzymes for milk processing, fruit juice clarification, meat tendering, and oligosaccharide synthesis...enzymes implies that even a minute quantity can significantly reduce the operational time for a given process... from their native conditions of ~0°C, which drastically affects their stable folding and efficient functioning...chaperones (the Cpn 10/60 isolated from Oleispira antarctica, and the P. haloplanktis Trigger Factor), that...Additionally, we’re also working towards widening the applicability of our strategy using the pMBP-LbCas12a and ...cultivating a nuanced understanding of the societal ramifications of laboratory science, and how it may best be...

...to the right indicates a higher signal, or upregulation, and a peak to the left indicates a lower signal...anything to the right of it, marking it as GFP+. Quantification for such graphs could simply be the percentage...case for you, switching to a contour plot should indicate where the two populations diverge. The smear ...place your gate based on the dot plot would be complicated.  Figure 2: (A) a smeary dot plot showing...Figure 4 compared to the dot plot in Figure 2. Quantification using MFI makes it easier to identify and study...Research Assistant in Arlene Sharpe’s lab at Harvard Medical School.   More resources on the Addgene blog Antibodies...addgene.org Addgene's Antibody Guide rAb Affinity Purification Protocol Video rAbs in the Addgene repository...

...emphasizing the importance of critical thinking, persistence, teamwork, and communication. And as a program focused...BioBuilderClub teams’ biological engineering designs and ideas ranged from a biological breathalyzer to pesticide... Dr. Natalie Kuldell from MIT’s Department of Biological Engineering. Built primarily from her 20 years...experiential learning. “Students may forget the technical details they hear in their classes,” she said,...and teachers to apply engineering principles to biological systems. Today, what started in Cambridge with...Apprenticeship, preparing local (Massachusetts), economically disadvantaged students for paid summer internships...

... microscopy. This application of AHA labeling is known as fluorescence noncanonical amino acid tagging...PhD students at the Max Planck Institute for Biophysical Chemistry, Göttingen in Germany. I attended this... symposium seeking inspiration to resolve the technical hurtle of monitoring protein turnover of specific...technique applies metabolic labeling using a noncanonical amino acid, azidohomoalanine (AHA). In the technique...DNA strands take part in rolling circle DNA amplification. The amplified DNA segment is labeled with fluorescent...directed toward AHA. Now the rolling circle amplification could only take place if the two antibodies ...interested in neurobiology.  She is passionate about communicating science to all audiences. Right now, she is ...

...and success. Customer and technical support Our customer and technical service brands continue to ...to keep up with the demand with excellence and dedication. We don’t have worry or wonder if providing...projects allows us to support more new projects. Diversification is happening. The Addgene team will have some... We score ~99.9% helpful and the customer and technical service teams put 100% effort into helping our...we might do. I am grateful every day for their dedication, flexibility, innovative thinking, and openness...Addgenies support a high collaboration, high communication, high efficiency, high quality, low meeting,...about the Addgenies. I never worry about their dedication, excellent intentions, or incredible drive to...

...red fluorescent proteins) allow simultaneous identification of more than one protein of interest. However... species (bottom panel in Fig. 1). Our results indicate that the tag protein may have an important influence...plasma membrane. Note that the use of the HA tag significantly improves the transport of hM4Di to the plasma...However, in some cases the fluorescent signal may be critical for the experiment. Since the fused protein could...fluorescence expressed in the neuropil conceals identification of opsin-expressing cells). For other things...Research Center. Using a combination of in vivo and anatomical techniques, her research aims to gain a deeper...Rodgers L, Wilson IA, Lerner RA, Wigler M (1988) Purification of a RAS-responsive adenylyl cyclase complex...

...repurpose popular GFP reporter lines for more sophisticated experimental manipulations, saving the time ..., 98% of TdTomato expressing cells were GFP+, indicating a robust, but specific system. T-DDOGs also successfully... types of proteins. They started with another biological workhorse, Cre recombinase, which induces recombination...systems are easily adaptable to neuroscience applications, including optogenetics, and they should make...Tang note that their scaffold system is likely applicable to many types of proteins. For proteins with ...Could Cas9-DOG be developed soon to allow genome modification only in a small subset of cells? Other proteins... PMCID: PMC4096992. Find plasmids from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “...