We narrowed to 626 results for: erf

...homemade competent cells of 107 CFU/μg are perfectly fine, yet performing multiple modifications will require... pathway to allow any cloning procedure to be performed using a simple two-step, 2 hr protocol prior to...adopted by any laboratory. All procedures are performed using the same three step protocol:  Fragment...efficiency of the strain: when asking the cell to perform multiple recombination events, all linear fragments... design (Figure 2), IVA cloning can be used to perform any plasmid modification, from insertions, deletions... reverse primer. Mutagenesis - Mutagenesis is performed by designing primers astride the old codon, amplifying...primers can be used in combination to simultaneously perform modifications, simply by adding all the necessary...

... line to perform downstream characterization of, isolating 2 or 3 distinct clones to perform key experiments...CRISPR-Cas system localizes to unintended sites and performs its programmed function there, this is an off-...off-target effects, but it’s important to not let ‘perfection’ get in the way of ‘good enough’. Predicting ...would confound your experimental results and 2) perform targeted off-target measurements at those sites... to quantify off-target effects entirely is to perform whole genome sequencing. While this may be unnecessary... The con to these approaches is that NHEJ must perform the repair to allow for detection, not Homologous...targets as well as clonal differences. If you are performing CRISPR library screens, the same principle holds...

...contrast technique in modern biology (1). When performed correctly, fluorescence microscopy provides a ...contrast image in which a bright signal overlays a perfectly black background. In addition to this, the use...confocal microscopes use parallelization to enhance performance. In spinning disk microscopy, a metal disk containing...coverslip at an angle such that when it reaches the interface between the glass coverslip and a sample in aqueous...within a few hundred nanometers of the glass/water interface. Super resolution microscopy Super Resolution ...means that the images will appear sharp without interference from blurry out-of-focus light. The most difficult...be needed. Most commonly these experiments are performed on a fluorescent widefield microscope. Here, excitation...

...extracellular fluid into the perfused liquid. The probe then collects the perfusion liquid for quantitation...fluid (Darvesh et al., 2011). This technique is performed with a probe surrounded by a semipermeable membrane... quantitation is desired (Figure 1). The probe perfuses liquid, which causes the passive diffusion of ...The advantage of this method is that it can be performed in a live awake animal by surgical implantation...into a specific brain region. While the method is powerful and offers immediate measurements in a live animal... GRABDA sensor here! Overall these sensors are powerful new developments in the detection and measurement...

...predict how fluorophores perform in ‘real experiments’, it is advisable to perform time-lapse imaging of ...To determine photostability, an experiment is performed that measures fluorescence intensity over time...Therefore, photostability measurements can be performed with soluble fluorescent proteins or localized...photostability measurements reported in the literature are performed in different ways. We describe a number of issues...fluorescent protein. Photostability measurements can be performed on purified fluorescent proteins. To avoid diffusion...changes in excitation power, it is important to perform the experiments on a single day on a stable system...

...donated MiSeq.  For Complete Plasmid Sequencing we perform a 2x251 run on our MiSeq that takes about two days...involves a significant amount of QC. First, we perform Picogreen quantification on each and every isolated...samples for Viral Genome Sequencing (VGS). QC is performed throughout the library preparation process and...Aligning the sequence The first step we take when performing quality control of newly deposited plasmids is...backbone sequence. We’re not necessarily expecting a perfect match - we will often find a few mismatches in ... reads to the correct sample. The PCR is then performed, optimized as needed, and cleaned up using a standard...to remove any template DNA and primers.  After performing extensive QC on that sample, we’ll then load ...

...for endogenous gene activation Optogenetics is a powerful tool that utilizes light to control and monitor... affinity for SspB1. In order to create a more powerful LID, the authors recently used computational protein...pathways. These LIDs are generalizable, versatile and powerful tools which allow you to easily and reversibly...elements in a single lentiviral vector provides a powerful tool to modify mammalian cells. Multicolor CRISPR...and broad host-tropism. These tools open up many powerful avenues through which one can examine molecular...all-optical electrophysiology is designing the perfect voltage indicator and optogenetic actuator pair...expression via western blot, to purify your POI, to perform pull down experiments like co-IPs and ChIP, and...

... with new techniques rapidly being developed. Performing a CRISPR experiment requires delivery of, at ...your genomic site of interest. This is commonly performed by transfecting cells with a plasmid, such as ...Cas9-gRNA ribonucleoproteins  An advantage of performing CRISPR experiments using RNPs is the variety ... RNP. This advantage has allowed scientists to perform in vitro and in vivo CRISPR studies in experimental...PEG-mediated methods, while technically less difficult to perform, often show low efficiency, while biolistic bombardment...plasmid based systems, most notably the ability to perform CRISPR studies in cells that are difficult to transfect...these have been acquired, genomic editing can be performed rapidly and, in many cases, with reduced chance...

...adjacent areas impacted the experiments we wanted to perform. It required a great deal of time and effort to... reformulating our AAV construct to optimize performance in our system and yet none of that data was published...packaging viral particles (6), can influence vector performance. Open science initiatives, like Addgene’s Data... to help scientists understand how our vectors perform in various experimental settings. It is a curated...low-magnification images demonstrating vector performance) will be assigned citable DOIs through DataCite... broadening the knowledge base of AAV vector performance in various brain regions and cell types. Power...potentially available to evaluate AAV vector performance. Yet, most of the technical data needed to replicate...

...applications. An antibody that performs well in immunohistochemistry may not perform well in a western blot. ... than mammalian cells, but mammalian cells can perform human-like post-translational modifications that... If that’s the case, a buffer exchange can be performed using a desalting column or an ultrafiltration...ones, consider increasing the number of washes performed during your harvest. Validation Recombinant antibodies... bioreactor supports high density HEK293 cell perfusion culture for the production of recombinant Erythropoietin...characterization & rheology, Current Opinion in Colloid & Interface Science, Volume 19, Issue 5, 2014, Pages 438-449...

...Created with BioRender.com.   Planning makes perfect Our first tip, and arguably the most important,...in silico cloning tools When available, always perform your MoClo assemblies in silico before you start...original Addgene kit plate for fresh bacteria. Performing your Assemblies Calculate your volumes As you...Save time, go high throughput You are ready to perform your assemblies, but you have a high number to ...to get through. It’s time to think big and perform your assemblies in a high-throughput format. If your...lowers experimental costs. If you are planning to perform a high number of assemblies, you will also likely...experienced MoClo user, but beginners or those performing complex assemblies should screen a minimum of...

...dTomato) that promote interactions between the FPs interfere with native interactions, preventing proper localization...cell-type dependent. Therefore, it is good practice to perform this assay with your chosen FP in your cell type...between studies is substantial (e.g, the poor performance of mRuby2 is detected by Constantini et al (2015...not by Cranfill et al (2016)). Moreover, good performance in the OSER assay is not a guarantee that using...relevant for your research. This will reveal the performance of fluorescent proteins in your biological system...resource with tools and data that indicate the performance of (a set of) fluorescent proteins under specific...

...are suitable for in vivo studies, our scientists perform all the standard quality control assays for AAVs... and Seqwell’s plexWell technology, we can now perform NGS at Addgene to sequence all the DNA packaged...should align to this reference sequence. Second, we perform a megaBLAST search on all the reads that did not...developed custom open-access Python scripts that can perform a deeper analysis of the data. Our serotype detection...expression. Next-generation sequencing (NGS) is a powerful tool that can be used to identify DNA contaminants...clinical-grade materials, and is not commonly performed for research-grade materials. Addgene is going...

... help take the pressure off having to find the perfect job immediately after you complete your Ph.D. or...to most postdocs. Commercial successes in the performing arts- not relevant at all. Please do not include...you are a classically trained ballerina who has performed at Lincoln Center. Your case should only focus...becoming the norm for EB-1A and EB-1B applications. Performing peer review at the request of your supervisor...counted as proof of judging. Merely being asked to perform is not enough. Casseus Law suggests that if you...sessions, panels, exhibit floor, etc.  These are wonderful opportunities to build relationships and connect...located in all 50 states and globally. Casseus Law performs in-depth vetting of a candidate’s background to...

...ecosystem Proven performance with other bioinformatics purposes  Integrated user interface, so our QC team...donation of a Illumina MiSeq sequencer enabled us to perform the necessary sequencing on-site, but now we were...already had a clear idea of the necessary steps to perform:      Fig. 2: Quick-stepping to a FASTA..., the overall purpose of a data pipeline is to perform several sequential computational tasks for which...smoothly operate the pipeline, we needed a user interface. In our next post, we’ll share our story about...

...eliminating the risk of the primary antibody interfering with downstream assays. This is particularly ... in the lysis buffer, keep samples on ice, and perform the IP at 4oC. When choosing the capture antibody... are completely removed. Wash steps should be performed even with a pre-cleared sample.  Eluting the target...and negative controls for an IP. When possible, perform the IP in parallel with both a sample known to ...order to identify any non-specific interactions, perform parallel IPs with beads only and an isotype control...affect the confirmation of the target protein or interfere with protein interactions of interest. Tagged ...

...reproducibility. Cause 1: Inability to repeat previously performed experiments Researchers should be able to follow...thorough notes on how lab procedures were actually performed, potentially curtailing future confusion (and ...previously published results. This over-reliance on the perfect story for a “high impact” publication can lead ...situation in Figure 1 where many scientists are performing similar experiments. Biology is messy enough ...practices that fall into this category include performing experiments with too few replicates leading to... an experimental design to a journal prior to performing the experiment (or set of experiments), the design...

...cells to form colonies on a plate and then we performed colony PCR on the register region and sent the..., each Sanger sequencing reaction can only be performed on one register at a time. If a particular application...considered as an alternative. The qPCR method can be performed on a sample of multiple registers. However, it...recognition site array on the register. RSM (See performance data from the indicated figures in our paper ... register. The purpose of this is to prevent interference between the transcribed gene and its 5’ UTR....have observed that certain GRSMs may still not perform as expected, perhaps due to idiosyncrasies that...

... for. When you complete each cycle, your best-performing mutant is usually the starting point for the ...choose, whether it's tweaking the antibody-antigen interface or fine-tuning the regions you want to evolve....increase an antibody’s binding specificity, you can perform binding assays against the target antigen to identify...temperature, can be adjusted to find the best-performing mutant antibody in those conditions. If you think...environmental conditions. These methods are especially powerful when coupled with iterative rounds of selection...selection and mutagenesis to refine antibody performance. If you are interested in learning more about antibody...

...Fluorescent proteins (FPs) offer scientists a simple yet powerful way to tag cellular proteins and investigate protein...scientists later developed red fluorescent PA-FPs by performing several rounds of random mutagenesis screening...most scientists working on in vivo imaging models perfer the PA-RFPs over PA-GFP. Another class of irreversible...fluorescent protein for your experiment Similar to performing experiments with standard FPs, before choosing...characteristics, as the monomeric state is less likely to interfere with endogenous protein structure and function...before your experiment even begins! If you are performing multicolor experiments, you will want to be sure...