We narrowed to 740 results for: NSI

...normally lies around 7.35-7.4 so mammalian cells are sensitive to pH. Anything between orange and light orange...Additional passages will allow your cells to achieve a consistent growth rate. For every minute spent organizing...

... 3-D path of the protein’s backbone in three-dimensional space, but their flexible and jiggly nature suggested...structure into experimentally determined electron density, have been known to say something silly like “See...

...nuclease-null—in order to effect different perturbations. Consider, for instance, the challenge of attempting to ...alterations can be exploited to quickly and inexpensively generate a potent Cas9-based transcriptional...

...can survive positive selection, but it will be sensitive to negative selection. Remove the positive selection...creating knockout animals or cell lines, please consider sharing them by depositing with Addgene! Figures...

...suppressors, and their protein products. Proto-oncogenes consist of a group of genes/proteins that, when expressed...Ian A., Paul D. Lewis, and Carla Mattos. "A comprehensive survey of Ras mutations in cancer." Cancer research...

... capture large field-of-views (6). For more comprehensive reviews on several other LSFM configurations...Bradke laboratories later developed the 3DISCO (3-dimensional imaging of solvent cleared organs) method of ...

...deposited by David Root of the Broad Institute) has consistently been one of the repository’s most frequently...Rory Macneil on Bitesize Bio Data Freedom: The Expansion of Data Sharing in Research Publications by Jim...

...students. But he also found that his teaching responsibilities left him with limited time for one-on-one ...notes the importance of acknowledging student apprehensions about “failure” (protocols not working, hypotheses...

...expression Whether BiFC constructs are expressed transiently by transfection or stably impacts the success...independent of PPIs. Simply compare the fluorescence intensity of your BiFC constructs to your negative control...

...enzymes do not contain the RuvC and HNH domains responsible for DNA cleavage, so they cannot directly edit.... PspCas13b does not require a PFS, and it is sensitive to mismatches in target RNA from bases 12-26 of...

...biologist--as an example of “big science,” a massive, expensive project that required collaboration from scientists...five Addgene colleagues who read it with me, the consistent response from scientist and non-scientist alike...

...For these reasons, we are in the process of transitioning our titering method from qPCR to ddPCR. Droplet...titer your AAV by ddPCR. Droplet digital PCR is a sensitive assay, so cross contamination into a no template...

...that is wide enough to dunk your plant’s flowers inside. 50 mL falcon tubes or plastic weigh boats are ...from a colony on a plate will reach an optimal density in 18-24 hours. I then add 0.5 mL of this starter...

...human CRISPRko sgRNA library The Brunello library consists of 77,441 sgRNAs with an average of 4 sgRNAs per...Mediator) uses an improved activator design that consists of inactive Cas9-VP64, a sgRNA with two MS2 RNA...

... Tris-buffered solution tends to produce more consistent readings — just be sure it is compatible with...these long enough?  There are, of course, more considerations and other approaches for measuring DNA quantity...

...category, will leave us with a single plasmid to consider. If you get this far and realize you need to apply...and let us help.  Check back on Thursday for an inside look at our mascot and homepage redesign. Additional...

...connections between biology and ancestral groups or, by extension, race. Many researchers do large-scale CRISPR ...potential for bias but to actively look for and consider bias as a confounding factor when interpreting...

...non-dividing cells. The first lentiviral vector system consisted of three plasmids: the packaging, envelope, and...capacity of a lentiviral vector but for which transient transgene expression is sufficient. Lentiviral...

...Saccharomyces cerevisiae in 1989 and are still used extensively to screen for molecular interactions in the cell...post-translationally modified, co-expression of the enzyme responsible for the modification in the assay host strain...

... Although we are busy with many other work responsibilities, we make team building activities a priority...at Addgene. In addition to her operational responsibilities, she enjoys organizing team building events...

...have, then bring them to your mentor. Pro tip! Consider physically walking through the protocol, by going...receive help until you’ve already started a time sensitive experiment. Ready to fly solo? Let your bench...

...with idea generation, try this thought exercise: Consider the issue as if there were no restraints on your...something completely unrelated to the issue and consider how it could be shaped to solve your problem. ...

...our own repository data, giving us significant insight into which vectors are likely to be of most use...Swanson, L. E., & Fan, M. (2016). Practical Considerations for Using Pooled Lentiviral CRISPR Libraries...

...plasmids tend to be high copy number. Here, we will consider the replication mechanism of plasmids under stringent...plasmid to balance the total plasmid number.  Let’s consider pSC101. This plasmid is stringently controlled...

...CRISPR/Cas Toolkit for plants includes 95 plasmids consisting of CRISPR/Cas nucleases, base editors, gRNA backbones...activation-based norepinephrine (GRABNE) sensors allow for sensitive and high spatiotemporal measurements of norepinephrine...

...partitioning system in the bacteria, but generally it consists of a centromere-like region on the plasmid, a ...2015) Imaging centromere-based incompatibilities: Insights into the mechanism of incompatibility mediated...

...editing field by making experimental workflows considerably easier, faster, and more efficient than previous...novice to expert-level researchers in achieving consistently successful CRISPR experiments. Jessica is a ...

...with BioRender.com. Sanger sequencing was considered the gold standard until about 2008 when a new...novo assembly within Geneious gives us additional insight into the assembly. Geneious will show how reads...

...from many other types of cancer. Mesothelioma is considered an orphan cancer; it is rare disease with only...non-resectable mesothelioma by the National Comprehensive Cancer Network, a coalition of the top 27 cancer...

...technical roles. Therefore, institutions are responsible for ensuring that either all affected scientists...and Brandeis universities have stated that they consider postdocs paid on stipends to be FLSA exempt, and...

...gain insight into protein structure and function (Pan et al., 2018). cDNA libraries, which consist of plasmids...

...d like to manipulate in mind. Now it’s time to consider where you would like to attack this gene. Should...sequences into HEK293T cells. The fluorescence intensity was classified into four groups (4; brighter than...

...transgene expression from split vectors has been inconsistent. A greater understanding of basic AAV biology...vector strategies and lead to greater and more consistent transgene expression. References: Allocca M...

...great model organism because they are easy and inexpensive to grow and maintain in the lab, have a short... worm, Caenorhabditis elegans, has been used extensively as a model organism since the 1970s when Sydney...

...this interaction will give you an idea of how responsive and supportive the ELN team is. Once you’ve made...information. If an employee leaves the institution, the transition is smooth as all data is organized within an ...

...have been described--CPEC (circular polymerase extension cloning) and SLiCE (Seamless Ligation Cloning ...multiple fragments, or synthetic biology, you should consider giving it a try! References: 1. Gibson DG, Young...

... Summers, 1998). Multimeric plasmids are also considered to be less stable and more prone to being lost...the supercoiled plasmid monomer, and at <6 kb, consistent with the size of a supercoiled plasmid dimer....

...into the mouse zygote (Qin et al., 2015).  When inside the zygotes, the gRNA will seek out its target ... Ong ST, Bergstrom DE (2018) Viable Mice with Extensive Gene Humanization (25-kbp) Created Using Embryonic...

... that have negative impacts on fitness. One consideration when making a transgenic cell line with the ...the piggyBac™ system, these challenges can be considerably reduced. The piggyBac™ transposon system can...

...training. Similarly, there are several factors to consider when selecting a postdoc mentor and lab. These... having sufficient data to write a manuscript. Consider questions like: What figures are missing? For ...

...regions you want to evolve. With this added layer of insight, you can shrink your mutant library and increase... Biotin. The antibodies developed by IPI are extensively characterized and validated, with the application...

...demonstrated that selection for repair of a temperature-sensitive pha-1 mutation significantly enriches for knock-in...from a single injected animal.  PAM sequence considerations For changes 5’ to the PAM, sequences corresponding...

...human proteins, this would be time-consuming, expensive, and not to mention frustrating. One elegant solution...transfection, add csRNP and ssODN to the cell suspension, transfect and let cells recover for 2-5 days.Note...

...successfully [10-12]. The next stage of reprogramming consists of redifferentiation of iPSCs into the cell type...pluripotent stem cells (iPSCs) by temperature-sensitive Sendai virus vectors. Proc Natl Acad Sci U S A...

...contacting [email protected]. These deposits are considered large deposits and are exceptions to our standard...and Christina Mork in February 2024.  Additional Inside Addgene Content on the Blog Quickest Way to Deposit...

...Looking for a fast and inexpensive way to determine your antibody's purity? Check out our latest protocol...

...updating our CRISPR guide. Our CRISPR guide is a comprehensive resource that covers most things CRISPR - from...

...in the scientific community. Addgene has been considering a gradual move off X as a response to major platform...

...Matthew Norton and NEBGCE4All: Making Genetic Code Expansion AccessibleResearcher: Ryan Mehl; Andy KarplusIBEX...

...itself. Many of us have experienced the minor yet consistent annoyance of walking into a fly lab and finding...