... procedure (Mortensen et al., 2019). Our method might provide some useful guidelines for the development...transformation was successful at all. Once we were on the right track, we switched to luciferases to more easily...simple and easy technique to implement. Success might vary between experimenters. Vacuum infiltration ...especially problematic with reporter genes as you might observe expression from Agrobacterium, but not the...leaf transformations, but we hope that these tips might help to get you started. Once you have established...

...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...

...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...

...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...

... 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...

...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...

... 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...

... 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...

...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...

...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...

...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!  ...