...plan and optimize your own protocols. Here we’ll start with just the first couple of steps - mainly, fixing...Fixation allows you to freeze your samples in a given state or point in time. You want to halt degradative processes... a protocol is available. When possible, try to start with conditions and reagents that have already been...for general IF protocols for your sample type. Starting with a protocol that usually works in your system...minimize the risk of it being lost from your sample, start by fixing with formaldehyde and permeabilizing with... Introduction to Performing Immunofluorescence Staining. Methods in Molecular Biology. Springer New York...

...methods.  In terms of preparation, VSV-G is stable and can withstand ultracentrifugation which helps scientists...alpha1-3)galactosyl (alphaGal) sugars were less stable than viruses produced in cell lines that do not... from gibbon ape leukemia virus (GALV) are more stable (Sandrin et al., 2002). Studying a viral pathogen... Q, Liu Q, Huang W, Li X, Wang Y (2017) Current status on the development of pseudoviruses for enveloped...modified RD114 envelope glycoprotein show increased stability in sera and augmented transduction of primary ...

...the conference is usually not enough. How do you stand out from the hundreds of others that did the same... Again, attending one of these fairs is a great start, but you need to do more than just show up. Do your...that you can tailor your CV accordingly. Make it stand out with key words and by putting your most relevant... particular employer in mind, you may consider starting with indeed.com or monster.com, where you’ll find... giving you a call. Many companies enlist the assistance of recruiters for identifying appropriate job...know very well; always ask for information and establish a connection with that person before ever asking...

... an enzymatically “active” state rather than reverting to an inactive state like Cas9 or Cpf1. As a result...homology to the crRNA or presence of a PFS. This is in stark contrast to Cas9, which requires that each DNA target... to activate programmed cell death or a dormant state for bacterial cells that have been infected with... we know about its structure and function? For starters, Cas13a can be used to bind and cleave target ...LwaCas13a), using a superfolder GFP fusion to stabilize the protein in mammalian cells. LwaCas13a-msfGFP...

...can have when starting in an academic lab. I floundered for a solid year after I started my postdoc! ...for scientists making this transition. In this installment of the Addgene career guide, we’ll think about...cloning and lentivirus production, I didn’t really start to recognize my transferable (or “soft”) skills ...you’re out of the lab to think about these--get started with our Transferable Skills Guide.     Over ...gloves again! The opportunities are there once you start looking for them. Acknowledgements Big thank you...

...depend on the host cell’s initiation proteins to start their own replication. In general, these plasmids...binding of the low-affinity versions is required to start the replication process. When enough copies of ATP-DnaA...replication and balance population growth against sustainability. Plasmid DNA replication and regulation Plasmids... the bacterial host's cell cycle, maintaining a stable concentration of plasmid. Similar to replication...near the ori and directly interacts with DnaA, stabilizing its binding and contributing to DNA unwinding...with a beneficial function (usually antibiotic resistance).  In the lab, low copy plasmids may result in...

...coli B strain, except ** which are K12 Strain Resistance Key Features Genotype Use BL21 (DE3)   Basic...pLysE (CamR) Expression of toxic proteins BL21 star (DE3)   Lacks functional RNaseE which results in...DE3) Δ(srl-recA)306::Tn10 (TetR) Expression of unstable proteins  HMS174 (DE3)** Rifampicin RecA-deficient... hsdR(rK12- mK12+) (DE3) (RifR) Expression of unstable proteins Tuner (DE3)   Contains mutated lac ... you wait too long to induce, your culture will start collecting dead cells, and, conversely, you cannot.... The pLys plasmid contains a chloramphenicol resistance cassette for positive selection and a p15A origin...

...applications. I think these papers are good starting points to understand the contours of each topic. The video...field is simply to know where to look and how to start. There is value, then, in having a guide that curates..., podcasts, and communities to help people get started in synthetic biology. While not everything is open-access...many people will find this guide useful as they start their synthetic biology journey. And if you have...

...designing entirely new proteins, improving our understanding of how protein folding works and creating biological...nature (at least that we’ve seen so far!). They started by designing a novel topology, then iteratively...expressed, purified, and characterized it, producing crystal structures demonstrating “the structure is strikingly...possible. Although Top7 was designed simply to be stable and novel, the principles that created it have ...novo designed proteins also helped researchers understand more about the biophysics of protein folding ...

...figurines that you can stage in a lab diorama. Or maybe you want to create a stack of wrapped gifts using...picture or video for either contest on Twitter or Instagram by December 31, 2021. Let us know which contest...and the winners will be revealed on Twitter and Instagram shortly after. Have fun, be creative, and enjoy...celebrates and decorates, follow us on Twitter and Instagram as we share pictures of our decked out spaces....

...assays using non-fluorescent visualization methods. Stay tuned for a future blog post to learn more about...experimental goals. Example steps include: counterstaining, mounting sample on slides, and confocal   .... To narrow down which specific methods to use, start by reading the methods sections of papers that have...straightforward than choosing a primary, so we’ll start there. Some of the most important considerations...commentary by Uhlen, et al. 2016 is a good place to start.) Reviewing available antibodies can seem overwhelming... Introduction to Performing Immunofluorescence Staining. In: Methods in Molecular Biology. Springer New...

...domain and the N-terminus of the other domain at a distance that favors proper folding and formation of the...tend to have lower affinities, lower long-term stability, and a higher likelihood to aggregate due to their...characterization.    Figure 2: Phage display starts with a pooled library of scFvs that are displayed...between the VL and VH domains. This bond provides stability and solubility for the scFv (Gaciarz and Ruddock...are inserted into a vector encoding the antibody constant domain scaffold to convert the scFv into an antibody...disulfide bond independent folding of intrinsically stable scFv. PLoS ONE 12:e0189964 . https://doi.org/10.1371...

...post was contributed by Shravanti Suresh from Iowa State University. Since its appearance, SARS-CoV-2 has...recommends one strategy: testing, tracking, and social distancing. With countries like South Korea, one of the ... cDNA. Isothermal amplification is done at one constant temperature, eliminating the need for thermocyclers...to our guest bloggers Shravanti Surest from Iowa State University. Shravanti Suresh is a PhD candidate ...CRISPR/Cas immune systems in the Sashital Lab, Iowa State University. She is actively interested in science... R. S., Baxter, N. J., Kosik, K. S., Arias, C., Acosta-Alvear, D., & Wilson, M. Z. (2020). A Scalable,...

...out there. The house mouse (Mus musculus) Let’s start with the most preferred mammalian model organism... set of mutations that made cancer cells more resistant to chemotherapy. You can find these plasmids at...found in lewy bodies), have been invaluable in understanding the basic molecular mechanisms underlying the...scientists can easily observe early developmental stages which can be difficult in other vertebrate species...assess developmental processes. For example, the Stainier lab studied the proliferation and differentiation...temporal expression of different transgenes, the Stainer group determined that there are actually two distinct...

... combination and possibly discover unexpected epistatic changes in protein activity. See Natarajan et ...product is digested with DpnI to remove residual starting template. After transformation and harvest, the... matter? Most of the time no, but there are 2 instances where it does. 1) When the backbone vector has...References 1. Natarajan, Chandrasekhar, et al. "Epistasis among adaptive mutations in deer mouse hemoglobin...Wrenbeck, Emily E., Justin R. Klesmith, James A. Stapleton, Adebola Adeniran, Keith E J Tyo, and Timothy ...  4. Fowler, Douglas M., Jason J. Stephany, and Stanley Fields. “Measuring the Activity of Protein Variants...

...editor installs the recombinase “landing sites,” then the recombinase handles the installation of the ...Free-floating rat brain sections stained with Anti-GFAP. The staining patterns from recombinant antibodies...methods such as chemical crosslinking or tilted-stage imaging. A truncated form of a tardigrade LEA, RvLEAMshort...the PASSIGE strategy. First, twin prime editing installs two sequences containing attB/P recombinase landing...

...tip boxes, and more (stay tuned for a few blog posts coming up about sustainability in the lab!). The @...red nosed reindeer.       View this post on Instagram     Eppi the red nosed Reindeer! 🦌🦌 as you.... If we've tagged you it's because we love your Insta!! #LILCChristmas . . #lab #ladiesinlabcoats #Christmas...phdtips #sciencecommunication #sciencedaily #instascience #ladies #research #cardiovascularresearch #parkinsonsdisease...

...my daughter has a business degree. From the very start of their undergraduate experiences, they were provided...begin to tackle these challenges, a group of stakeholders came together with the goal of driving systemic...scientific training and employment infrastructure. A Stakeholder Workshop at Janelia Research Campus in 2019 solidified...Evidence-Based Models for STEM PhD Career Development; a Stakeholder Workshop Report.” The report enabled the development... Additional resources on the Addgene blog Get started with Addgene's science career resources Browse ...

...for that. Lately, significant advances in the understanding and application of CRISPR/Cas9 technology are...news was circulating. Scientists at North Carolina State, in the labs of Chase L. Beisel and Rodolphe Barrangou...still working out important details in their understanding of how CRISPR/Cas9s actually work. Last month...of Cas9's three-dimensional shape via x-ray crystallography, an advance they say should allow scientists... report showing Cas9's structural details. "Understanding this structure may help us engineer around the...

...readers about how our articles have helped them understand different topics of science, especially disease...sessions led by scientists, which is hosted on our Instagram channel every Sunday. What's planned for the upcoming...scientists and non-scientists. We’re in the process of starting a forum where non-scientists can directly approach...participate in our one-hour Live Q&A session on Instagram every Sunday. Undergraduate and graduate students...scientific jargons. Someone should also be able to understand what you are trying to convey without having ...