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We narrowed to 1,005 results for: RING

Showing: 261 - 280 of 1005 results

  1. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    ...wash buffer such as PBS or TBST. Stringent stripping The stringent stripping solution uses heat and detergent...your sample? Or what if something weird happened during your western and your results look… funky? Do you...along with the antibodies, especially with more stringent stripping methods. If you are planning to detect...that section of the blot unusable for reprobing. Preparing your western blot Run your gel and membrane transfer...primary and secondary antibodies from your membrane, clearing the way for the second blot. There are multiple...with a mild stripping solution and move onto a stringent solution if the mild is not effective. You may...perhaps using longer incubation times, or try the stringent solution if you started with the mild one. Reprobe...

  2. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...of choice is AAV, with nearly every experiment requiring AAV. Before joining my lab, I had never worked... a heated stir plate to dissolve and requires monitoring so that the solution doesn’t get too hot because.... Rather than investing all of this effort in preparing reagents, I choose to only harvest the cell pellet...layered and then the viral suspension generated during AAV production is overlaid. Following ultracentrifugation...first purification. This helped me get a feel for layering the iodixanol solutions and let me double-check...make one or two small aliquots to use for qPCR titering. This helps avoid multiple freeze/thaws of your...of virus. AAV titration by qPCR Overview: AAV titering by qPCR quantifies the number of genome-containing...

  3. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ... steps, until its connection with DNA is broken during cross-link reversal (not shown in diagram). Cross-links...sequence targeted in the experiment. Figure 2: Comparing CAPTURE and CAPTURE 2.0.     Original CAPTURE...) one or more gRNAs Two separate lentiviruses delivering 1) a BioTAP-tagged dCas9 and an eGFP tag, and...detecting chromatin interactions opens the door for answering more questions about chromatin conformations. ...‘social networks’ of 157 super enhancers.  3. Measuring temporal changes in chromatin interactions A third...in chromatin conformation, like those that occur during development and cellular differentiation. Using...the Xu Lab tracked enhancer-promoter interactions during erythroid differentiation. CAPTURE 2.0 captures...

  4. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...packaged inside the vectors during production. These impurities cannot be removed during the purification step...recombinase-independent recombination during plasmid growth in bacteria or during viral production. This recombination... control assays for AAVs. These steps include measuring viral titer and assessing purity. But Addgene ...wrong plasmid? What if bacteria snuck into the prep during the production process? These questions are especially...materials. Addgene is going the extra mile and pioneering the systematic use of this new QC assay to guarantee...Evidence for Encapsidation of Prokaryotic Sequences during Recombinant Adeno-Associated Virus Production and...

  5. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ...path began to take shape, was when I started volunteering. I wasn’t sure yet what a career in science ...volunteered at a place that made me happy. I started volunteering in the “Dinosphere” almost every week. I gained...dinosaurs, and met wonderful people. When I started volunteering, I was worried about becoming one of those angry...t sure what I wanted to do. A few weeks into volunteering, I was talking to a little girl about fossils...Beyond the outreach experience I gained from volunteering, it gave me a place to go once a week to get...before founding CINSO. Now, the network I built during graduate school is my informal advisory board. ...to share valuable life lessons they have learned during their scientific training and wrote about them ...

  6. Which Fluorescence Microscopy Techniques is Best for Me?

    Type
    Blog Post
    ...the palm of your hand. Although white light is entering your hand, only orange/red light can be seen exiting...optical section is achieved that boosts contrast by filtering out emission light from fluorophores higher in...as spinning disk confocal. However, due to the scattering nature of biological tissue, imaging more than...parallelization (7) have allowed for real-time monitoring of neuronal activity across substantial 3D volumes... allow for multi-view imaging can image thick scattering samples in their entirety, something that is ...excitation light which can lead to photobleaching when acquiring multiple images over a large axial range. 2-photon...overcome this (8). The recent development of tissue clearing techniques (9) now allows researchers to image...

  7. Tips for arabidopsis transformation

    Type
    Blog Post
    ...they begin flowering. It is important to perform the transformation not long after flowering begins. If...RFP in the seed (Emami, Yee and Dinneny, 2013). Preparing plants for transformation Whatever plants you ...you can cut off the primary inflorescence (the flowering portion of the plant) near the base of the stem...al., 2006. Tips: Don’t plate seeds too densely during selection: this will increase background growth... (2002) Arabidopsis : a laboratory manual. Cold Spring Harbor Laboratory Press. Available at: https://...). Additional resources on the Addgene blog Engineering the plant genome using CRISPR/Cas9 Using pSiM24...pSiM24 for simplifying plant genetic engineering Using arabidopsis in the classroom Resources on Addgene.org...

  8. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ...system to partition themselves to each daughter cell during cell division. They don’t simply rely on chance...CBP), and a partition NTPase (Schumacher, 2012).  During cell division, CBPs bind to the centromere-like...replication that occurs for some types low copy plasmids during cell division simply does not allow for enough ...strategy has been successful in displacing, or “curing” the virulence plasmids from the bacterial pathogens... SR, Iredell JR (2017) Plasmid interference for curing antibiotic resistance plasmids in vivo. PLOS ONE...X, Wang D, Wang H, Feng E, Zhu L, Wang H (2012) Curing of Plasmid pXO1 from Bacillus anthracis Using Plasmid...22253811 Ni B, Du Z, Guo Z, Zhang Y, Yang R (2008) Curing of four different plasmids in Yersinia pestis using...

  9. Viral Production at Addgene

    Type
    Blog Post
    ...customers. Details about our production protocols, titering methods, and quality control are described below... or through collaboration with viral vector manufacturing facilities, such as the University of Pennsylvania...are then aliquoted and stored at -80 °C. Titer Titering is either performed by Addgene or by the University...University of Pennsylvania Vector Core. In general, titering is performed by the facility that produced the...those achieved using the standard qPCR method.  Titering by the University of Pennsylvania Vector Core ...Purity Purity of AAV preparations is assayed by comparing the relative stoichiometric ratios of the viral...aliquoted, frozen, and stored at -80 °C. Titer All titering is performed on lentiviral preparations that have...

  10. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    ...appeal, instant sharing across multiple other sites), I created a new platform for sharing video abstracts...University and developer of wesharescience.org. Sharing your research with the world can be challenging...printed journal articles as the dominant tool for sharing ideas. This leads to everyday obstacles for many...www.WeShareScience.org. WeShareScience is a free platform for sharing short, 3 to 5 minute videos about research. Videos...oceanographer who has a blog and several videos on sharing your research through video. Liz Haswell and ... 525.7569 (2015): 306-307. 3. Spicer, Scott. "Exploring video abstracts in science journals: An overview...Scientific Impact Data Freedom: The Expansion of Data Sharing in Research Publications Resources at Addgene....

  11. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...the questions and preparing answers early in the day. Although I didn’t start answering until 5pm, frankly...well moderated. One of our original worries about sharing science advice on Reddit was that we’d be dragged... our AMA and Joanne’s preferred time to start answering questions. r/Science has pretty strict rules about...straight on through her designated two hours for answering questions. She was able to do this because we ...posted at 8:00 am and Joanne didn’t have to start answering until 5 pm). The ability to see the questions ... Tips: Online Social Networking for Scientists Sharing Your Lab Protocols: Using Apps to Save Time & Track...Experiments Additional Resources See How We're Sharing Science through Video with the Addgene Youtube ...

  12. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...culture has become overly confluent or too old. Ensuring that you are working with healthy, happy cells... the lentiviral stock needs to be titrated. If storing for less than a day, lentivirus can be kept at ...dry ice/ethanol bath or liquid nitrogen prior to storing. Know your target cells Labs tend to follow the...titer the virus (read this blog post for tips on titering your lentiviral prep!) and infect. While this ...consider is how the virus is being titered. Most titering approaches transduce into highly permissive cell...receptors, other important binding events take place during the course of infection. These binding events can...23589850. 9. Cronin, J., Zhang, X.Y., Reiser, J. “Altering the tropism of lentiviral vectors through pseudotyping...

  13. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...clicking “Add Plasmid to this Article” (left) and entering a plasmid name,plasmid type, and plasmid description...the plasmid names and descriptions first before entering the data. This step associates an Addgene ID with...plasmids all built with the same backbone and only differing in the gene/insert. Pooled libraries are normally... should click the "Enter Data" button to begin entering the specifics of your plasmids. Figure 5: Click... plasmid information. Enter plasmid information During the remainder of the process, you will enter plasmid.... You can complete these pages sequentially by entering the appropriate data and pressing "Save and Continue...species that the gene or insert sequence came from. Entering this information will populate a list of possible...

  14. Celebrating Outstanding CRISPR/Cas9 Achievements at the Dr. Paul Janssen Award Dinner

    Type
    Blog Post
    ...acceptance addresses were inspiring! We also got to hear more from them during a panel discussion with ...bestowed upon discovers of CRISPR/Cas9 genome engineering, Dr. Jennifer Doudna and Dr. Emmanuelle Charpentier...the presence of scientific heroines is always inspiring. I was especially excited to attend this event...enough to make up for the somewhat naïve questions. During the panel, an artist was sketching her interpretation...accelerate the launch of clinical applications. During the panel discussion, Dr. Mello took it upon himself... in the room. He offered the great analogy of comparing the CRISPR/Cas9 system to Norton Antivirus. Antivirus...

  15. London Calling - Addgene’s New Europe Office in London

    Type
    Blog Post
    ...of Plasmid Sharing: Interested in more statistics from Addgene's 10 years of plasmid sharing? Check out...our US counterparts are still sleeping). Plasmid sharing in Europe: Addgene’s request statistics Since 2004...Developmental Physiology at the University of Paris VII. During my years in research, I developed several new plasmids...is excited to be working at Addgene, because it brings scientists together from all over the world to ...discussing a new scientific discovery or invention, and sharing those ideas with others. This love of interacting... blog post Or look at our "10 Years of Plasmid Sharing" Infographic!  ...

  16. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ... recently showed that by delivering CRISPR components as RNPs and delivering two halves of a repair template... Precision Medicine, CRISPR, and Genome Engineering. Springer, Cham, 2017. 29-42. PubMed PMID: 29130152... used AAV gene editing to introduce naturally occurring cancer-causing mutations into the endogenous allele...homology arms with the modification of interest, ordering them from a gene synthesis company such as IDT...anaphase, and chromosome segregation in human cells harboring tumor-derived mutations in STAG2." PLoS genetics...Howes, Rob, and Christine Schofield. "Genome engineering using adeno-associated virus (AAV)." Chromosomal...

  17. Transferable Skills Guide: Time Management

    Type
    Blog Post
    ...goals are (see monitoring task progress), and I know that I’ve blocked out time during my day according...transferable skills while working in the lab and by offering advice on areas where you might need some help... can spend accomplishing small tasks (like... preparing plasmids to send to Addgene). Following my advice...too many activities too close to one another. Monitoring task progress. For this, I always make sure I...-world example, let’s say I’ve blocked out time during my day to write a blog post, edit other blog posts...leave your phone in your bag on “Do Not Disturb” during the times you’ve blocked out for other activities...instance, while I may have found splitting cells boring, hanging out in the tissue culture room also gave...

  18. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...structure determination LEA proteins protect samples during cryo-EM grid freezing Engineered and evolved optimizations...2024.01.21.576499.    LEA proteins protect fragile samples during cryo-EM grid plunge freezing By Rachel Leeson Cryo-EM...can be damaged at the air-water interface (AWI) during the freezing process, resulting in sample loss....protective methods or detergents like CHAPSO, while requiring a lower concentration of sample — nearly ten times...-water interface protectant for fragile samples during cryo-EM grid plunge freezing. bioRxiv 2024.02.06.579238...prime editing systems for specific applications! Delivering prime editors (PEs) via engineered virus-like...off-target editing. Cargo size limits are also less stringent for eVLPs than other viral delivery methods, allowing...

  19. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ... for monitoring RTK signaling.  Figure 4: Design of pYtag fluorescent biosensors for monitoring RTK activation...for investigating neuronal signaling and for engineering improved sensors for these and other targets....class of signaling receptors, but the options for measuring activity of specific RTKs are limited. In a new...colleagues describe pYtags, a modular system for monitoring specific RTK activation in vivo (Farahani et ...sensors. Using genome mining and structure-guided engineering of potassium binding proteins, they developed...and are a promising starting point for further engineering. Figure 5: Structure-guided design and genome...s41467-022-30685-x. Feel the heat with B-gTEMP  Measuring local temperature variations and heat transfer...

  20. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...of animals. There are many different naturally occurring strains of E. coli, some of which are deadly to...strain is a bit more convoluted due to researchers sharing and renaming samples throughout history. It was...cloning, routine cloning, cloning unstable DNA, preparing unmethylated DNA, and more. Many mutations that...Consequence dam DNA adenine methylase mutation (GATC) Preparing unmethylated DNA, important when trying to cut... dcm DNA cytosine methylase mutation (CCWGG) Preparing unmethylated DNA, important when trying to cut...relA1 Relaxed phenotype, mutation eliminating stringent factor Allows RNA synthesis in absence of protein...lacZΔM15] hsdR17(rK-mK+) JM110  Streptomycin  For storing plasmids that should not be Dam or Dcm methylated...
Showing: 261 - 280 of 1005 results