...budding. Yeast Centromere plasmids (YCp): These are considered low copy vectors and incorporate part of an ARS...are most similar to bacterial plasmids and are considered “high copy”. A fragment from the 2 micron circle...spontaneously occuring resistant mutants and the insensitivity of yeast strains to some antibiotics. Auxotrophy...uses for said element. This link provides a more extensive list of yeast auxotrophic markers and includes...auxotrophic selection in E. coli? Additional considerations HIS3 L-hisitidine no S. cerevisiae  no yes...ade6+ purine (adenine)   S. pombe  no no   Considerations when using auxotrophic selection Of course,...

...interested in following? Someone you admire for their insight and way of expressing it in meetings? The more ...help you learn the local “ropes” and become an insider faster A mentor that works elsewhere is able to... If your organization has a mentoring program, consider tapping into it and helping make it work by being... coaches who charge for their advice and time. Consider this option but be cautious and make sure to choose... to be your mentor, it may sound like a big responsibility or commitment. Start by asking them to have...questions and topics to discuss. The mentee is responsible for leading the discussion and for being “coachable...

...recurrence, and treatment of cancer have been extensively explored. Biologists have attempted to study ...and dynamic phenomenon that is interestingly responsible for tumor reduction as well as progression. Dunn...is a region of incessant chemical activity, and consists of other cell types, especially immune cells and...relapse. Chemical and radiation therapies cause a transient shutdown of the immune system, hampering its ability...have been made to use whole cell therapeutics consisting of various types of immune cells. Dendritic cells...exhibit cytotoxic activity. CAR-T cells have consistently shown promise in reducing leukemia and lymphoma...impacted by the trial. During clinical trials, expansion of participants based on early results can enhance...

...thin plane within a relatively thicker three dimensional sample (see Figure 1); similar to an MRI or CT...rapid photobleaching and a reduction in signal intensity. 2-Photon microscopes restrict excitation (and...focal point of the objective can a a high enough density of red-photons be establish for this to happen....field of the objective (the distance in the axial dimension that will appear in focus). This means that the... then collected and projected back to a fast, sensitive detector such as a scientific CMOS camera. If ...Reversible Saturable Optical Linear Fluorescence Transitions (RESOLFT/STED) also excel on these types of samples...a host of tools and opened doors to many great insights, it can be overwhelming to a novice microscopist...

...in vivo, they have higher reproducibility and consistency in the final product. Because the constant (non-epitope...binding) region is the first thing to go when downsizing an affinity reagent, most of the alternative ...its thorns and every affinity reagent has its downsides. These reduced-in-size options often have reduced... as ankyrin repeat proteins. DARPins typically consist of 3-4 repeats, have high specificity and binding...it? This post isn’t meant to convert anyone to downsize their affinity reagents, trendy as that may sound...Grumezescu, A. M. (2018). Drug targeting and stimuli sensitive drug delivery systems. Elsevier Science & Technology...

...LoxP vector.   Turn it off for a little while Transient suppression of expression is faster and easier...molecules that degrade target mRNA transcripts. When transiently introduced, they can deplete endogenous protein...instead of introducing a DNA double strand break. Transient introduction of CRISPRi or inducible expression...expression. Addgene has a dCas9 KRAB plasmid for transient knockouts!  Turn it on Turn it on forever Permanently...activator and repressor mechanisms Want to transiently induce expression? Check out Addgene's Lentiviral...site-specifically knocked in. This can be labor intensive and time consuming. If your native protein doesn...

...experimental design considerations used in all behavioral studies. General considerations for behavioral ...behavioral testing As a general consideration for behavioral testing in a laboratory, a high level of experimental...increase stress levels and affect behavior. A fact to consider: Rodents can hear ultrasonic frequencies (> 20...targeted to a specific area. Some other technical considerations for stereotaxic injections are: Vector titer...brain areas, repeated staggered injections may be considered. Timing: High expression levels of the viral ...

...isotype choice. Target and application should be considered The first question you may have is whether a ...monoclonal primary antibody (A). To boost signal, consider using subclass specific secondary antibodies instead...immunoprecipitations. Your sample species should also be considered when choosing an antibody. In some applications...options are available to you, it may be worth considering an antibody raised in a different species.    ...      Table 1: Isotype considerations when choosing antibodies   Complexity is a key factor...with a distinctly conjugated secondary antibody. Consider too that some secondary antibodies can cross react...

...Your time is precious. Here are a few things to consider before you commit: Double check to make sure ...can be a particularly good resource if you are considering using a reagent for “off-label” purposes. When...about using Google for serious science questions, consider using Google Scholar, which searches scientific...email you’re writing is probably not directly responsible for the problem. We’re here to help. You've ... few days without getting a response, however, consider following up. Ask for a timeline if you are ...upfront how long it will take to get an answer, consider asking to avoid unrealistic expectations. And...

...details on how they work!) Figure 1: Base transitions are edits between the purine bases (adenine, ...limitation to this approach. CBEs and ABEs are base transition editors. They adjust the side groups of DNA bases...task, not a job for a single enzyme. Base transitions Base transversions C → TG → A A → GT → C ...some cases, the same repair pathways that were considered problematic in the development of CBEs and ABEs.... Like other ABEs, the editor they were using consisted of Cas9 fused to an engineered adenosine deaminase...rapidly. If it follows a similar path to base transition editing, we can expect several generations of...to repair DSBs. Base Editing terms   Base transition Base changes between purines (A ↔ G) or between...

...infects host cells through binding the human angiotensin converting enzyme 2 (ACE2) cell-surface receptor...receptor. The spike protein RBD responsible for binding ACE2 is the target of most anti-SARS-CoV-2 antibodies..., homogeneous time resolved fluorescence, and suspension bead assay (Figure 2). After finding no cross-reactivity...scFVs to cognate aaRS antigens as measured by suspension bead assay. Image from  Preger et al., 2020. ... a compact Cas9 ortholog from Staphylococcus lugdunensis that recognizes a NNGG PAM and has comparable...expressing GCaMP6f with this promoter. Image calcium transients in PV interneurons with AAV-S5E2-GCaMP6f are ...

...solution against a part of the body very sensitive to heat- the inside of the wrist or the cheek. (Parents ...work with, require extra serum to survive at low densities - adding to the expense of dilution cloning. ... transfected cells into 3 plates at different densities for selection). A 2% agarose solution is then ...plates containing selection agent at different densities, then dispose of the plates that do not appear...such as the Viaflo from Integra Biosciences) to transition from a 48-well cell plate to a 96-well assay ...

...class of FPs, the FP timers, add an entire new dimension to monitoring localization; using FP timers, researchers...production of dsRed E5. This timer predictably transitions from green to red fluorescence (500 to 580 nm...However, this initial timer was a tetramer with a propensity to aggregate in cells (Tsuboi et al.), limiting... in Xenopus embryos. Of course, timers may be sensitive to changes in oxygen, temperature, and pH, but...et al. developed monomeric FP timers with less propensity to aggregate. These monomeric FP timers (mK-GO... that differentiates human IPSCs into glucose-sensitive insulin-secreting beta-like cells." Nature communications...

...is its simplicity and versatility. CRISPR/Cas9 consists of two components: the non-specific endonuclease...libraries (referred to simply as “CRISPR libraries”) consist of a heterogeneous population of gRNA-containing... is right for you There are several factors to consider when selecting a CRISPR library for your experiments...identified using next-generation sequencing (E). Considerations and tips for successful screens Next-generation...check out Addgene’s 2016 publication “Practical Considerations for Using Pooled Lentiviral CRISPR Libraries...Waxmonsky NC, Swanson LE, Fan M (2016) Practical Considerations for Using Pooled Lentiviral CRISPR Libraries...

...As a person with many roles and responsibilities, I have to think a lot about how to balance those roles...meeting and a means to request funding for events, consider reaching out to it. PTAs have more flexibility...schooler himself and b) I had the background to be considered an authority in this area. What is genome engineering...presentation, she said “it was such a valuable extension to our curriculum and offered engaging examples...Educational Progress (NAEP), virtually none were considered proficient in science, regardless of race or ...posts Resources on Addgene.org Check out our comprehensive CRISPR guide Find CRISPR plasmids and resources...

... blog post discusses some of the key things to consider when performing colony PCR.   Designing colony...for the presence of many different inserts. The downside: this type of primer does not provide information...information about insert orientation, then you might consider designing orientation-specific primers. Blunt ...day, you can save the leftover bacteria-water suspension and use them to start cultures of your positive...up PCR reactions: The remaining bacteria-water suspension will serve as the template for your PCR reaction...tool to have in your molecular biology tool kit. Consider giving it a try next time you’re screening for...

...provenance (this is the history of the cell line consisting of records of the origin and propagation of the...cell line contamination: Epithelial Mesenchymal Transition Is Required for Acquisition of Anoikis Resistance... The same test is so good that it is used as a forensic technique (aka DNA fingerprinting) that can place...convince people of some of the basic tenets of responsible cell culture outlined in this article. Now many...studied biochemistry at Warwick University, gaining extensive cell culture research experience through working...for a range of testing, from animal doping to forensics, DNA sequencing to drug testing. You can find ...

...cells, thus expressing additional EBNA-1 transiently. The transiently expressed EBNA-1 increases protein expression...field [4]. Three major reprogramming methods are considered when generating hiPSCs for clinical purposes:...purposes: mRNA transfection- this allows transient expression of the reprogramming factors Sendai virus- a non-integrating...efficiently [5]. However, the Sendai virus method is expensive, especially if used to make clinical grade hiPSCs...OCT3/4 and shRNA against p53 pCXWB-EBNA1 37624 transient EBNA-1   Many thanks to our guest blogger,...

... elaborated on later. As for what job fairs to consider, some of the best options are the Naturejobs Career...most relevant experience at the top. You may also consider visiting any of the employer’s career pages to... apply for the position. This post on Business Insider nicely summarizes good job fair practice. Career...’t have a particular employer in mind, you may consider starting with indeed.com or monster.com, where... important to realize that you have to take responsibility for your own professional development, not ... Reading and Outside Resources http://www.businessinsider.com/how-to-get-the-most-out-of-job-fairs-2014...

...includes the aforementioned internal control and is consistent with the result in Figure 1 could look like this...associated with the independent variable should be consistent over time and from different preparations of ...replicate control is important, it speaks to the consistency of the reagents and hands performing the experiment...experiment, not the reproducibility or consistency of the observed phenotype associated with the independent...replicates control for the reproducibility and consistency of the observed phenotype associated with the...may be, proper controls are a basic tenet of responsible scientific inquiry and investigation. References...