...found that the accuracy of ICE analysis results was highly comparable (within r2=0.96) to that of NGS data...coefficient (r) is also computed and reported.  The higher the R2 value, the more confident you can be in ...the Download Analysis Data button on the bottom right of the analysis screen. ICE analysis details Contributions...genomes that contain the indel. The discordance plot (right panel) shows the level of disagreement between the...that continues after the cut site, representing a high level of sequence difference. Traces The Traces...

...scientists achieve a high titer (reviewed in Joglekar and Sandoval, 2017). This high titer is one of the...your virus, there are many other reasons when you might pseudotype your virus:  Lowering cytotoxicity. ...reaching the target cells. VSV-G is also toxic at high concentrations as described above and is inactivated...vector to be transported retrograde and also has higher neuronal transduction efficiency and a better safety...

... develop and optimize the method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK... the promiscuous cleavage and degradation of neighboring ssRNA and ssDNA by Cas13 and Cas12a, respectively... of CRISPR-based diagnostics. SHERLOCK- Specific High Sensitivity Enzymatic Reporter UnLOCKING Cas13 (...SHERLOCKv2 provides a method to detect nucleic acids with high sensitivity and specificity without compromising...the ability to rapidly detect nucleic acids with high selectivity and sensitivity from patient samples...

...technology has become very popular as a rapid and highly efficient way to move DNA sequences into multiple...two new recombination sites (attL-left- and attR-right-, Figure 1). Under certain conditions, the attL ...’ll walk through an example experiment where we might use Gateway cloning to generate our desired constructs...another, the inserted DNA fragment stays in frame. High efficiency The positive (antibiotic) and negative...M. (1992). A Genetic Switch: Phage (Lambda) and Higher Organisms (Cambridge, MA: Cell Press). Additional...

...three scenarios, but a short treatment regiment might wipe out bacteria that are completely susceptible...concentrations of a drug, but will be killed by a higher concentration, or sometimes more exposure time ... corresponds to the level of susceptibility you might expect if a certain antibiotic were chosen for treatment...survive for millennia. What we can do, however, is fight smarter, not harder. We can utilize tactics like...antimicrobials to learn how scientists are using CRISPR to fight bacteria. From a societal standpoint, we can all...

...Department of Labor’s general guide to the effect on higher education, you can see that those in teaching roles...lot of us, frankly quite surprising. Consortia of higher education institutions submitted letters to the...exemption. Discussions about the drastic effects on higher ed had been seized upon by opponents of the ruling...convincing as the recommendations for postdocs to be on higher salaries (which have been made by multiple organizations...recommendations that postdoc salaries be raised to $50,000 - higher than the level they will be raised to on Dec 1st...

...you can see some of the stats from the AMA to the right - the Reddit community is active, engaged, and has...resources at her disposal, she was writing all day and straight on through her designated two hours for answering... to other scientists who will find it useful, I highly recommend doing an AMA! Before I turn it over to... Kudos to the moderators for keeping the quality high and thus enabling a productive discourse. They even...to resources. I’ve said this before, but I also highly recommend taking every opportunity you get to speak...

...cells. However, for some applications, integration might be considered a double-edged sword with unintended... lentiviral vectors, how they work, and why you might choose them over integrating lentiviral vectors....lentiviral vectors? There are several reasons why you might choose an IDLV over a lentiviral vector: 1. No risk...mind is that titering these IDLVs may require a slightly different protocol than regular lentiviral vectors...Reyon, D., Joung, J. K., & Sander, J. D. (2013). High-frequency off-target mutagenesis induced by CRISPR-Cas...

...created vectors for Lambda Red that contained one of eight bacterial toxin genes isolated from E. coli or Psuedomonas...both rhamnose- and arabinose-inducible promoters. Tight control of toxin expression is absolutely necessary...induction system is well-characterized and quite tight (Giacalone et al., 2006). The Chen lab demonstrated... selected the three toxins so that at least one might be useful in a broad array of pathogenic Proteobacteria...expression in Escherichia coli using a rhamnose-based tightly regulated and tunable promoter system." Biotechniques...

... cell mixes, like bulk lympho-/splenocytes, you might only be interested in one of the various cell populations...markers of all cell types except CD8+ T cells. We might choose the markers CD4, CD19, and CD11c, which would...rest can be excluded, or “dumped”. From left to right: B cell, CD4+ T cell, dendritic cell, CD8+ T cell...host species reactivity. When sorting cells with a high resolution for specific downstream applications,...immunity. When he's not in the lab, Paul enjoys weight lifting, sailing, and reading.   More resources...

... a great plan to start consistently knocking out high-quality videos for you all. The next day...we got...Everything we had set up involved multiple people in tight locations. Conference rooms for on-screen presenters...raised on an ironing board, giving it the perfect height (Fig. 2). We recorded video using a simple, external...comfortable on camera while thinking about things like lighting, camera angle, and shot composition.  So back ...could transmit a signal between a wall. There was a SLIGHT delay, but I could work with that.  Focus: Next...

...change and screen a few hundred clones to find the right one Step 2 – Edit the cells a second time using ...corrects the blocking mutation and again identify the right clone from a few hundred. From these two steps, ... few hundred clones - Still better than 10.000, right? (Kwart, Paquet, et al  2017). Find Additional CRISPR...increasing mutation incorporation efficiency, we might exploit this relationship to direct the zygosity...Additional Resources on the Addgene Blog Learn about High Fidelity Cas9 Variants How to Design Your gRNA Delivering...

... together on a tight budget. To help popularize our kit resources,  we’re highlighting just a few of ...covering a wide range of fields from Addgene. What you might not know is that Addgene distributes curated collections...Morgan’s blog post on the PUF Assembly Kit for some insights on its potential from its creators. We hope you...

...amount of high-quality DNA, real-time PCR has several advantages including easier automation, higher throughput...method called pyrosequencing. The drive for faster, higher throughput and less expensive sequencing fuels ...commercially available although they are reported to have high error rates (10). It is projected that nanopore-...and research progress. These will bring faster, higher throughput, and potentially more accurate techniques...

...untreated individuals. Symptoms of AIDS include rapid weight loss and an increased risk of infection, including...models, and it’s difficult to tell if one approach might be more successful. “Shock and kill” has the advantage...expression of Cas9/gRNAs, which could result in higher rates of off-target cleavage. In both cases, translating...each of these gRNAs will need to be validated for high on-target binding and low off-target binding. It...stable expression of Cas9/gRNA. Viral production is higher in this model than in latent HIV-1 infection, which...

...the perfect empty backbone, made sure to add the right tags to your gene, and may have even put a fluorescent...are marketed for a specific purpose: fast growth, high-throughput cloning, routine cloning, cloning unstable...  Propagation of ccdB-containing plasmids Hte   High transformation efficiency deoR constitutive expression...synthesis Allows uptake of large plasmids hee   High electroporation efficiency supE44 (glnV44)   Suppression...+)  XL10 Gold Tetracycline and Chloramphenicol High competency cloning and propagation of large plasmids...

...This Depositor Week, we're highlighting eight amazing posts on research done in our depositor labs. Some... Andrew Willoughby introduced us to RUBY-red, a bright red pigment reporter for plants.  Cas7-11 and PASTE...

...community like Watertown to bulk purchase energy from a higher percentage of renewable sources. This program offers...sources and we have chosen to participate at the highest level offered. As a result, we will be drawing ... our electricity from renewable energy projects right here in New England- the majority of which are solar...temperatures, biosafety cabinet ventilation, and lab lighting/HVAC requires a significant energy expenditure...

...Sleigh bells, please! It's time to announce the winners of the 2022 #DeckTheLab contest!  and it's.....in our building. Unfortunately, snow is thawing right now, so today noone could see my (not so) little...                     A post shared by 草方 (@gracie_fang) That’s right, researchers of the world. Cap your Eppendorf tubes...Instagram! Happy decorating, all, and to all a good light!    🤩 our lab in Bayliss Building @SMS_UWA is getting...

...vector pAAV-CAG-RdLight1 AAV9 Biosensor Tian New viral vector pAAV-hSynapsin1-RdLight1 AAV9 Biosensor...lab. Also of interest is the Tian lab's pAAV_hSynapsin_psychLight2 (AAV9), which contains a biosensor that...Biosensor Tian New viral vector pAAV_hSynapsin_psychLight2 AAV9 Biosensor Tian New viral vector pAAV.hSynapsin.SF-iGluSnFR.A184S...

...unless the source of culture is irreplaceable or highly expensive). Detecting mycoplasma contamination ...induces a noticeable color change. This method has high sensitivity but low specificity (Degeling, 2012)...required in this process because result interpretation might be difficult. The low density of mycoplasmas, contamination...media as much as possible and the ones that are tightly packed with seals. Use disposable pipettes to avoid...exposure of the cell culture to air and make sure to tighten the flask cap before placing them in the incubators...

...decades of my life following a very straightforward conveyor belt from high school to college to graduate school...Veterans Health Administration, I could never get one straight answer about what science policy even is! In this... “little p” policy.  “Big P” policy is what you might typically think of. It occurs in a government setting... challenging if you are trying to apply for an outright job in a field where you have no specific scientific...

...that infectious wild-type viruses were produced in high-titer retroviral stocks generated by the transfer...In this kind of approach, a population of cells might express only the tTA or your transgene resulting...developed, but these vectors are quite large and you might get lower viral titer when using them.   Regarding.... If you are transducing non-adherent cells you might need to centrifuge your cells with the virus in ...Trouble Shooting Viral Transductions Learn Why You Might Want to Use AAV in Your Research Lentiviral Vector...

...and four chaperones from Denis-Yarmouth Regional High School visited our labs in Cambridge, MA to learn... Outreach Scientist Jessica Welch showed some highlights of his travels from 2015, including a great European...Website Learn more about Dennis-Yarmouth Regional High School Resources at Addgene  Read more Inside ...

...working with AAV or lentivirus in your research, you might want to use one of our virus backgrounds. Addgene...backgrounds for you as well! If you use FRET, you might be interested in our fluorescent protein background....           Did you get the joke? If not, you might want to check out Addgene’s Fluorescent Proteins...background.     We hope these science backgrounds will brighten up your virtual meetings!  ...

...It’s well known that exercise can help fight stress and depression, and now researchers have reported...including unpredictable loud noises and flashing lights - that sent average, control mice into depression...know about you, but I think I could use a good run right about now. And, if you are looking to get your hands...for studies in your own lab, you’ve come to the right place. Get started with PGC-1 plasmids from Addgene...

...Directors brings human resources experience and insight to the now 13 member board at Addgene. Let’s meet...hope Addgene will accomplish in the near future? Right now it’s a matter of maneuvering through the impact...volunteer appointments included the Belmont Human Rights Commission and mediator for the Massachusetts small... dedicated to volunteerism and ran the Mayor’s Midnight Sun Marathon in Anchorage, for the Leukemia Society...

...to the field. Fortunately, our bodies have been fighting cancer for far longer than we’ve been studying...natural abilities, we’re gaining new ground in the fight against cancer in its many forms. The dichotomous...cancer stops growing, and the immune system stops fighting. Such quiescent cancer may rear its head once ...infection can further divert the immune system from fighting cancer. Surgical removal causes increases in tumor... a sequence of immune reactions that aid in the fight against cancer. Sipuleucel-T by the Dendreon Corporation...

...instance, you might use it as part of a flex switch you’d like to work with in neurons or you might use it in...Cre-controlled gene of interest makes pmSyn-BFP-Cre highly modular. Further uses of pmSyn1-EBFP-Cre Of course...brain. There are many possibilities enabled by this highly modular AAV - that’s probably why it’s so popular...

...bigly. But such a project should not be undertaken lightly. Whether executed in arrayed or pooled format there...are just as critical for success. Sometimes, you might have a few positive control genes on which to test...execute genome-wide perturbations. So, while some might argue that the only way to do something is to do...still offering the potential for true biological insight. Pooled and individual plasmid formats A first ...