We narrowed to 740 results for: NSI

...Kebabian, John W., and Paul Greengard. "Dopamine-sensitive adenyl cyclase: possible role in synaptic transmission...Gary L. Petzold, and Paul Greengard. "Dopamine-sensitive adenylate cyclase in caudate nucleus of rat brain...

...equipment needed for EM or SRM? Enter Expansion Microscopy. What is Expansion Microscopy?  ExM provides a straightforward...done about that.  Second, expansion introduces distortions. The goal in expansion is for your sample to expand...maximizing label density. In some cases, nanobodies have been found to help improve label density (Mikhaylova... Bournonville L, et al (2021) Expansion microscopy provides new insights into the cytoskeleton of malaria...Expansion Microscopy (ExM) promises an easier, more accessible way to image biological features previously... ExM images in a new publication.         Why Expansion Microscopy?  Traditional light microscopes are...samples can be expanded to ~4-10x linearly! This expansion physically separates molecules while maintaining...

...investigator who is using an in vivo animal model should consider the possible elicited immune response from viral...

...was aware that such roles existed, but did not consider them for myself. I started picturing myself in...technical support specialist for 4 years. Read on for insights from my time in technical support including tips...takes some time (several years) to build up comprehensive lab experience. With this valuable training ...any incoming requests or problems - there is considerable diversity in the types of questions customers...individual stories and get (sometimes refreshing) insights into how other scientists think and work. Most...drive to help others. These roles offer an easy transition from an academic lab to industry without being...

...serve Addgene and ourselves. We are also able to consider different technical approaches that may come up...

...s post, we’ll discuss some of the important considerations you should think about when developing optogenetics...place aversion also models reinforcement but is considerably simpler than ICSS. The test chamber is divided... of the type of behavior, certain technical considerations still apply. Testing stimulation First, In ...cue. The hippocampus has long been known to be responsible for mediating this behavioral effect but the ...mouse; however, all the steps and technical considerations are directly applicable to the rat [6]. Technical...PMC3150193. 8.Han, Xue, et al. "A high-light sensitivity optical neural silencer: development and application...

...Plasmid Sharing Or read more about what happens Inside Addgene   ...

...development of transient transformation systems for other species as well. Things to consider in developing...generating a transgenic plant can take even longer, a transient plant leaf transformation process could save the...transgenic plant takes even longer, is more labor intensive, inefficient, and often not reproducible. Prior...to creating transgenic cultures or plants, our transient expression method enables us to quickly screen...biosynthetic pathway. Why plant scientists do transient leaf transformations The development of transgenic...common problem in non-model organisms. Therefore, transient methods are usually the first choice for investigating...instead of months to years. In the plant sciences “transient transformation” usual means that neither a stable...

...may be time, then, to look towards genetic code expansion (GCE): techniques in which non-canonical amino...introduced to the bacteria using plasmids. Rapid expansion occurred, with hundreds of new ncAAs and techniques...number of experiments with. The catch The rapid expansion of GCE led to the creation of many new tools and...meaning that the final protocol may involve a sensitive balance of trade-offs. This balance can be extremely...Tan, Z., & Cornish, V. W. (2021). Genetic Code Expansion: A Brief History and Perspective. Biochemistry...

...Understanding the transition While a PhD equips you with exceptional technical skills, transitioning into management...   Strategies for a successful transition To successfully transition into a managerial role, it's essential...mentors who have made similar transitions can provide you with insights, lessons from their experiences... most pivotal decisions many PhDs face when transitioning from academia to industry is whether to stay...preparation and a willingness to embrace new responsibilities. After my second postdoc, I realized that...career trajectories within the industry, but the transition wasn’t easy - I had to reinvent myself and redefine...d like to share below. Common pitfalls in the transition (what I wish I knew before experimenting on myself...

... as channelrhodopsins (ChRs) makes them light-responsive. When light shines on excitatory channelrhodopsin-expressing...tissue culture dish but can also happen in real-time inside a mouse’s brain.  But the brain is a tricky organ...mm3). Current channelrhodopsins also need high-intensity light for activation which requires implantation...design new channelrhodopsins that are more light sensitive and have enhanced conductance compared to current...channelrhodopsin’s photocurrent strength, wavelength sensitivity, and off-kinetics.   This model was then used... measured photocurrent amplitude, wavelength sensitivity, and off-kinetics correlated with the model’s... produced stronger photocurrents at low light intensity than existing channelrhodopsins CoChR and/or ChR2...

...should be an important consideration during any experiment. Sensitivity Cas9 sensitivity is often closely related... important factors to consider: site accessibility, specificity, and sensitivity. Site accessibility Site...Another important consideration when choosing a Cas9 is balancing specificity and sensitivity. Wild-type Cas9..., accessibility, specificity, and sensitivity are key considerations in choosing a Cas protein for your...appropriate for your specific research question. Considerations when choosing your Cas nuclease When choosing... related to specificity. Sensitivity is a measure of the on-target activity of the Cas9 nuclease. This... measure of off-target activity (left) while sensitivity is the measure of on-target activity (right)....

...technique! What is a photosensitizer? First, for a few definitions. A photosensitizer is a dye or protein...system? Turns out that with light-inducible photosensitizers and a quick zap of the proper color light,...light, you can do just that.  Photosensitizing dyes and proteins have been around for awhile (check out this...Bruchez and Tsang labs recently developed a photosensitizer composed of the protein complex complex and... oxygen species (ROS). The ROS produced by photosensitizers inactivate target proteins or cells while ...having little effect on neighboring cells. The photosensitizer used in this work is composed of two parts:...fluorogen (the TAP - targeted and activated photosensitizer - MG-2I in this case). The fluorogen is not...

...to individual researchers. Yet accurate and comprehensible organization of data is essential for efficient...store raw data in formats that require specific (expensive) software that evolves over time such that prior... the journals themselves should be primarily responsible for storage.  If datasets are too large to transfer...projection of it, could allow idea generation and extension, based on existing, published data. The importance...However, expectations must also be reasonable, sensitive to the data type and made as facile for both the...

...and quantify based on band intensities of your DNA relative to the intensity in the ladder. It’s best to...things to consider: cost, time, equipment, and expected DNA concentration. It’s important to consider the advantages...any special reagents. However, there’s limited sensitivity at low concentrations of DNA and it can’t distinguish...measure all nucleic acids). These methods are more sensitive than UV absorbance, especially when you expect... to compare the band intensity of the fragment in the ladder that is closest in size to your piece of ...nm. This method is time consuming and has low sensitivity so it’s not used much. However, the measurements...

...and a Spanish minor, it’s time for me to start considering my future path. Talking to Addgenies has helped...biggest takeaways from my co-op. It has been so insightful to learn about different career paths, successes...

...researchers, we wondered how we could make the insights gained during this time more visible and thereby... ReFigure saves time, knowledge and makes your insights discoverable. Watch this quick video to learn ... ground again and again as they don’t save the insights they glean from a reading session. To remember...notes on a “Pinterest for scientists”? ReFigure extension is a simple Chrome plugin that allows you to save...papers to find the right experiment! The ReFigure extension also tells readers if a paper they’re reading ...ReFigure making it easy for others to discover your insights on a topic. Beyond collating other’s data, you...instantly and are editable by the author at any time. Insights contained in your ReFigure could be beneficial...

...networks, and specific neuronal populations to consider. To tease apart how different neurons impact a...experimental approach you choose, it’s essential to consider the technical limitations of each approach and...

...conducted in Professor Cepko's lab and to get some insight into her management and mentoring styles. If you'd...

...better and make scientists’ lives easier. Why transition to Addgene? So why did I join Addgene? Was I ...easier. I’m hoping my story demonstrates that transitioning from academia doesn’t have to mean abandoning...graduate training. PhDs have many roles to play inside, on the outskirts, and entirely outside of academia...

...chemicals, and are not always consistent. Precast gels, while expensive, reduce protocol time, can be ...chemical-based blockers, while much more expensive, are consistent and stable at room temperature for 1–2... Direct detection method Less sensitive; usually more expensive Nitrocellulose or low-fluorescence...Multiplex Fluorescent proteins Less sensitive; more expensive Strip/Reprobe PVDF membrane Increased...specificity, lower cross-reactivity, and are more consistent lot-to-lot. The highest reproducibility comes...quantitative assays. On the other hand, they are not as sensitive to lower amounts of protein, and can require more...  Lysis buffer When choosing your lysis buffer, consider the subcellular location of your protein. Proteins...

...antibody is less sensitive than HRP RANbody detection, it is approximately as sensitive as detecting the...detection is less sensitive than HRP RANbody detection but approximately as sensitive as antigen detection...approximately as sensitive as detecting antigen with a standard primary/labeled secondary Less sensitive than HRP-RANbody...approximately as sensitive as detecting antigen with standard primary/labeled secondary   Less sensitive than HRP-RANbody...antibody and a ≥100-fold increase in signal over intrinsic fluorescence. Increased signal is likely due to...secondary antibody. Chicken IgY This RANbody reporter consists of two chicken IgY domains. Chicken IgY domains...detected enzymatically with HRP substrate More sensitive than detecting antigen with a primary/ labeled...

...      Figure 1: A protein gradient. The density of the bands, which increase as protein concentration... buffers and strengths. If that’s not working, consider using a subcellular fractionation kit or a lysing...re seeing protein aggregation, you may want to consider changing your incubation temperature. Some proteins...aggregate at 95 °C, a common lysis temperature, so consider a longer incubation (10–20 minutes) at 70 °C, ...If you need very accurate protein measurements, consider using an unstained protein ladder, which will ...ladder on the lefthand side. Whatever you do, be consistent and document it.   Did it work? Save yourself...reversible staining method of choice is appropriately sensitive. If using a PVDF membrane, you can dry the membrane...

...detailed below. Voltage Sensitive Domain (VSD) GEVIs Simply put, VSD-based sensors consist of a VSD fused to...gated ion channels all depend on transmitting and sensing electrical potential. Thankfully, there is a tool... These tools have two main elements: a voltage-sensing component and a fluorescent output. The GEVI family...divided up structurally into 2 main groups: voltage sensitive domains and Rhodopsin containing GEVIs. Chemigenetic...fluorescent reporter and the mechanism of their voltage sensing component, which separates them into two distinct...indicator, which can make the kinetics of voltage sensing to signal output slightly slower than other approaches...approaches. Functionally, the voltage sensitivity of the available VSDs has been successful even at the...

...the loading control: the intensity of each POI band is divided by the intensity of the loading control ...band intensity measurements become extremely susceptible to error, since tiny changes in intensity can ...membrane, which would be directly reported by band intensity. Researchers use later steps in this chain (blue...trust Assumption #1, any change in the normalized intensity of your POI could reflect either a genuine change...relationship between protein abundance and band intensity is an S-shaped curve. Sparse proteins may not ...problems for proteins of very different abundance, consider the following example gel. Figure 5: An... on the plot of protein abundance versus band intensity (right). Created with BioRender.com. On this...

...Specific ​H​igh ​s​ensitivity ​E​nzymatic ​R​eporter un​LOCK​ing) technique Sensitivity: Single molecule...Two-step-SHERLOCK has a sensitivity in the zeptomolar range while One-step-SHERLOCK has a sensitivity in the femto-aptomolar...for rapid, ultrasensitive and visual detection of novel coronavirus SARS-CoV-2 Sensitivity: 4.6-11 copies...SARS-CoV-2 using a CRISPR based DETECTR assay Sensitivity:10-100 copies of RNA per microliter Time: ~40...two crRNAs that bind to two regions improves sensitivity of the assay. This reaction also contains SSB...one-temperature reaction, and is nearly single-molecule sensitive. In the case of HIV-1 and SARS-CoV-2 detection...detection of SARS-CoV-2 genetic material (CREST) Sensitivity: 10 copies of RNA per microliter Time: Not mentioned...

...conditions. The use of low intensity light solves each of these problems; inexpensive tools such as LEDs can...precise, consistent, and controllable pulses of light. Complicated patterns of light intensity and wavelength... low-intensity light, so off-target effects should be limited. Phytochromes are light-responsive systems...Tabor have been working on making E. coli light responsive since 2005, when Tabor was working in Voigt's... heading of two-component systems (TCSs). TCSs consist of a histidine kinase that phosphorylates a response...Christopher Voigt’s lab created the first E. coli light-sensitive two-component system (TCS). Levskaya et al. fused...Tabor et al. subsequently engineered a second photosensitive TCS activated by green light; these two systems...

...published Aug 30, 2018 and updated April 16, 2020. Sensitive and specific nucleic acid detection is crucial...nucleic acid detection however, either lack the sensitivity or the specificity to detect nucleic acids at...at low concentrations and/or are too expensive, time-consuming, and complex to use outside of standard...and Cas12a, to develop simple, portable, and inexpensive platforms to reliably detect nucleic acids at...and optimize the method termed Specific High Sensitivity Enzymatic Reporter UnLOCKING (SHERLOCK and SHERLOCKv2...CRISPR-based diagnostics. SHERLOCK- Specific High Sensitivity Enzymatic Reporter UnLOCKING Cas13 (C2c2) was... your targeted nucleic acid. To increase the sensitivity of the assay, targeted DNA or RNA from a sample...

...are not as sensitive, however, so those studying low abundance antigens may want to consider alternative... ELISAs for their accuracy, specificity, and sensitivity (a good ELISA can detect picogram quantities ...advantages and disadvantages of each, and some considerations when using the method. ELISA basics All ELISAs...detectable product. Scientists compare the signal intensity of the sample to that of a standard curve to extrapolate... themselves, the process can be laborious and expensive. In some cases, conjugation methods can interfere...Incorporating a secondary antibody increases the sensitivity of the assay, since multiple secondary antibodies...sandwich ELISAs, in a competitive ELISA the signal intensity is inversely proportional to the amount of antigen...

...retain fluorescent intensity during exposure to light, is another factor to consider. If your application...interiors to many inquisitive scientists. Seeing inside these specimens has offered illumination on biochemical...will outline some fundamental characteristics to consider when selecting a dye or ligand to make your selection...proteins are SNAP-tag® and HaloTag®. These tags consist of a genetically encoded enzyme variant that covalently... is bound through green light emission. Key considerations in selecting fluorescent dyes/ligands Effects...-conjugation affects protein function and its propensity to bind cellular components. Some dyes target...stably combined with nanoparticles, maintaining intensity at ~85% of its original value after 60 minutes...

...subcloning, expansion, and confirmation. This procedure can take several months but should be considered in labs...benefits and drawbacks of different methods (for comprehensive protocols for all of the methods discussed here...results in less than a day, but tends to be less expensive than an ELISA kit. In this approach, viral RNA...lentiviruses in parallel, this approach should be considered. Similar to the p24 assay, titering via measurement...When using qPCR based methods for viral titration consider using a reverse transcriptase mutant in a separate...carryover. Functional titer Functional titer is considered a more accurate method to quantify virus as it...of this approach is that it can be quite labor intensive. Target cells are transduced with serial dilutions...

...the least expensive of all the options discussed as it only requires PCR and the inexpensive DpnI enzyme...here to help with that! Four important factors to consider for any experiment are: Speed Cost Difficulty...For cloning in particular, another important consideration is: The number of inserts ...as some methods...the following cloning methodologies have been extensively described in other articles; so if you are interested...little as 30 minutes! Alternatively, you might consider Gibson cloning. In this technique, you first perform...order determined by their homology. You might also consider FastCloning, first described by Chaokun Li and...Finally, TOPO cloning is another good option to consider when time is of the essence. This method requires...

...cellular-level events and behavior. It’s a problem that he considers “one of the greatest challenges of neuroscience...particular advantage for observing events taking place inside cells that are on the move, such as T lymphocytes... calcium-binding sites as possible for added sensitivity. As reported in Nature Methods, the FRET changes...essentially all of the standard organisms for which extensive genetic tools are available. While he recognizes... he said. “It gives you more information than intensity-based readouts.” Griesbeck also points out that...approach he and his colleagues took to optimize sensitivities can now be applied to dozens of other published...for visualizing the transcription factor cAMP-responsive element-binding protein (CREB) in living cells...

...classes of optogenetic tools, and what factors to consider when choosing one.     Figure 1: Example...or inhibit specific neurons without needing to consider secondary signaling pathways. There are many different...different opsin (natural and engineered proteins responsive to light in combination with retinal) variations...reviewed in Abreu & Levitz, 2020), including light-sensitive chimeric GPCRS (OptoXRs) (Tichy et al., 2019),... the extracellular domain of endogenous light-sensitive receptors, like mammalian rhodopsin, and the intracellular...Created with BioRender.com.   Important considerations for using optogenetic tools Once you’ve selected...specific tool. There are many, many options to consider, and the following factors will help you determine...

...your plasmid yield is sub-optimal? If you are consistently getting sub-optimal plasmid yields from your...Addgene uses and recommends DH5𝝰! This strain consistently gives good, high quality preps due to its endA1... this handy guide from OpenWetWare for a more extensive listing of common E. coli strains. Increase the...media or adding supplements can improve your cell density, leading to an improved plasmid DNA yield. Culture...supplements to the media You can achieve higher cell density using a nutrient-rich broth such as Terrific Broth...in moderation) may also serve to increase cell density and plasmid yield. Start with a single, fresh colony...Increasing the incubation time can improve the cell density and therefore the plasmid yield. You can optimize...

... One consists of NLuc fused to enhanced green fluorescent protein (GpNluc) and the other consists of NLuc... (FRET) probes. These probes generally consist of a responsive protein (e.g. calcium binding protein) ...membrane-inserted voltage sensing domain (LOTUS V; Luminescent Optical Tool for Universal Sensing of Voltage). With...how bright you can make a reaction but for how sensitive you can make a reaction. NLuc requires 100-fold...) fused to two fluorescent proteins. When the responsive protein binds to calcium it changes conformation.... In well-designed FRET-based biosensors, the intensity of the FRET signal can be correlated to specific...No photoexcitation High Ion specificity High sensitivity Easy expression Gives ratiometric data The result...

...each event to a channel number based on intensity. Higher intensity events are assigned to higher channels...a user will compare the shifting intensity of events. Since intensity positively correlates with expression...cabinet with cloning rings or following labor-intensive limiting dilution protocols? If this sounds like... the cell’s size, complexity and fluorescence intensity.   What flow cytometers detect Cell size and... a high degree of internal complexity such as extensive membranes produces a greater SSC.  Protein expression...The electrical system converts the fluorescence intensity to a voltage pulse, called an event, and assigns...reporter on the N- or C-terminus of the protein. One downside of fusion proteins is that the tag can alter the...

... to be considered as well. In this post, we’ll touch on antibody storage, buffer considerations, and give...records, and consider the following questions as you move forward:  Was a light-sensitive conjugate on...you’re planning to run with the antibodies time-sensitive, using precious samples, or being compared to ...data in a manner that demands high accuracy and consistency?  If the answer to any of these questions is...saving the poor, abandoned antibody for a…less sensitive moment.   Fig 1: Blugene fading into darkness...fix it. HRP is easy to find, simple to use, not sensitive to light in its inactive state, and cheap. Unless... panel in the machine. Fluorophores are light sensitive, so antibodies conjugated to them need to be stored...

...more sensitive channelrhodopsin actuator for triggering action potentials at low light intensities, the...cumbersome. To overcome many of the limitations intrinsic to DNA-guided systems such as TAL effectors or...antibodies fused to sfGFP. This system amplifies the intensity of fluorescence signal and enables tracking of... H+ specific response cytoplasmic retention insensitivity to other ions In BRET, A mutant Renilla luciferase...dimers by fusing the SsrA peptide with the light-sensitive LOV2 domain of Avena Sativa phototropin 1 (AsLOV2...activator protein to drive transcription. The anchor consists of dCas9 fused to CIB1 and is targeted to specific... a photoswitchable fluorescent protein which transitions from emitting orange (λ= 565nm) to far-red (λ...

...fluorescence intensity remains. The first issue is that, in several cases, the fluorescence intensity decay ..., fluorescent proteins may display reversible intensity changes (Shaner et al, 2008; Bindels et al, 2017...experiment is performed that measures fluorescence intensity over time. In order to predict how fluorophores...proteins and by comparing how the fluorescence intensity changes over time, the photostability of the fluorescent...photobleaching rate does not depend on fluorescence intensity or protein distribution. Therefore, photostability...analogous to radioactive decay). Consequently, the intensity decay is often described by the t½, which is the... it is necessary to know how the fluorescence intensity evolves over time. The second issue is that, in...

...help with transitioning out of academia (because so many scientists are making this transition). Another...have experience and knowledge to help scientists transition out of the academic sphere into a different meaningful...not always as effective in mentoring for this transition. Second, a coach will help you set goals and ...successful outcome and the good ones take this responsibility seriously. As I make my rounds speaking at ...However most coaches who focus on scientists are sensitive to this and have coaching plans that are financially...here are a number I have come across for you to consider. Feel free to add other suggestions in the comments...scientific backgrounds. For a stated purpose of transitioning a scientist from academia to industry, you might...

...2024.12.20.629623.  A fast and responsive voltage indicator with enhanced sensitivity for unitary synaptic events.... Z. (2024). A fast and responsive voltage indicator with enhanced sensitivity for unitary synaptic events... tags for single-molecule biophysics Fast and responsive voltage indicator ONE-GO biosensors kit Antibodies...collaborators recently created ASAP5, a faster and more responsive genetically-encoded voltage indicator (GEVI) ...ASAP5 selected for its improved kinetics and responsivity in screening ASAP3 variants. B) Electrical and...regimes with fast activation kinetics and high responsivity near the resting membrane potential, ASAP5 is...the formation of Gα-GTP by BRET. The biosensor consists of a YFP-tagged G protein and a NanoLuc® (Nluc...

... problem. It is estimated that mycoplasma is responsible for up to 60% of the cell culture contamination...contamination (Uphoff, 2002). Mycoplasmas are considered to be one of the simplest and smallest bacteria. The absence... source of culture is irreplaceable or highly expensive). Detecting mycoplasma contamination Detection...Drexler, 2011). Culturing on agar plates/broth is considered the ‘gold standard’ for detecting mycoplasmas...noticeable color change. This method has high sensitivity but low specificity (Degeling, 2012). Bioluminescence...result interpretation might be difficult. The low density of mycoplasmas, contamination by other bacteria...you keep all the required equipment and reagents inside the laminar airflow before and while you are working...

...Gervais C, Grazzini E, Durocher Y. Transient Gene Expression in Suspension HEK293-EBNA1 Cells. Methods Mol...based on magnetic beads from very high cell density suspension. Biotechnol Bioeng. 2021 Sep;118(9):3499-...At its core, harvesting recombinant antibodies consists of separating antibodies from media, cells, and...this method, beads are incubated with the cell suspension. Antibodies bind ligands on the beads and a magnet...alongside your ladder and a protein standard. The density of the antibody bands can be measured against the...multiple contaminating bands or a few strong ones, consider increasing the number of washes performed during...conformational changes in the antigen or lower detection sensitivity of the assay. Where possible, it can be useful...

...system both favors antibiotic sensitive bacteria and prevents sensitive bacteria from acquiring resistance...system. These CRISPR antimicrobials successfully resensitized a beta-lactam resistant population to the antibiotic... may result in plasmid loss and antibiotic resensitization, or may also cause cell death. Image from Beisel...et al. designed a two-phage CRISPR system to resensitize antibiotic-resistant bacteria (Figure 2). The...Bacteria that receive this phage are now antibiotic sensitive, but lytic phage-resistant. When the lytic phage...killed, antibiotics can be used to target the sensitive population. Yosef et al.’s proof-of-concept work....   Figure 2: Two-phage strategies to resensitize bacteria to antibiotics. A lysogenic phage infects...

...in yeast with Rosella, a pH-sensitive fluorescent biosensor A pH-sensitive fluorescent biosensor-based...-emission biosensor Rosella consists of DsRed connected to a pH-sensitive variant of GFP (SEP) by a 9 ...Virus (WNV) induced cell death. The Wu lab library consists of 77,406 individual gRNAs targeting a total of...Moduclar Cloning (CIDAR MoClo) Parts Kit. This Kit consists of a variety of plasmids containing many different...plasmids to the scientific community. This collection consists of plasmids containing genes from a crowd-sourced...scientists use GMAP, there will be a continuous expansion of compatible backbones, promoters, and genes ...all under the control of the cI857 temperature-sensitive repressor. The temperature-controlled expression...

...also incorporated LOV-Turbo into TransitID experiments.  Find TransitID plasmids here! These two developments...genetic cargo Optogenetic proximity labeling and TransitID Tracking cellular history with protein ticker ...37612276.   Optogenetic proximity labeling and TransitID By Mike Lacy Proximity labeling can identify protein...utility of this technique.  By engineering a light-sensitive LOV domain into the TurboID enzyme, Lee and Cheah...plasmids here! Next, Qin and Cheah et al. described TransitID, where two proximity labeling enzymes, TurboID...catalyzing the biotinylation of nearby proteins. B) TransitID uses sequential labeling by TurboID and APEX2 ...trafficking within and between living cells by TransitID. Cell, 186(15), 3307–3324.e30. doi: 10.1016/j....

...Cell lines should be routinely subcultured at densities that are low enough to prevent confluency but ... in lower lentiviral titers. Another factor to consider is the age of your culture post thaw. In general...can be subtle and may include reduced saturation density, decreased rates of growth, or clumping. To ensure... and resource consumption due to the high cell density of the culture. In addition to benefitting your...lower titer virus. In such cases you may want to consider concentrating your viral prep. Storage can also... start to a successful infection, one point to consider is how the virus is being titered. Most titering... that routinely produce lentivirus may want to consider testing a variety of cation and serum sources ...

...experiments a bit more challenging. With these considerations in mind, when planning an experiment users ...transgene, and downstream applications should all be considered when choosing the correct system. Recently, ...volume. Consequently, you will end up with a considerably lower volume than with the unconcentrated prep...experiment requires a certain volume of virus, consider scaling up by either increasing the number of ...while the PEG precipitation method is relatively inexpensive and does not require an ultracentrifuge, PEG ...lentivirus, it is time to titer. One important consideration when titering a lentivirus is the cell line ...transducing a target cell line. One should also consider the specifics of the titering transduction experiment...