We narrowed to 740 results for: NSI

...Cas enzymes and variants available. Without a comprehensive resource, it can be overwhelming to choose a...resources throughout the site. In the Experimental Considerations section on each enzyme page, you'll find a ...

... they flank the gene of interest. A single ITR consists of palindromic arms (A-A’, B-B’, and C-C’) that... Deng, J., Hebben, M., & Liao, J. (2024). A Comprehensive Study of the Effects by Sequence Truncation ...

...integrated into the genome of the target cell or transiently expressed. It’s important to note that for all...vector can express your construct in target cells transiently or constitutively depending on the virus used...

...Addgene also has a number of insect vectors to consider. An easy way to see what is available is to browse...transgenic fly There are a few important points to consider when choosing the vector you’d like to use to ...

...described in this publication We also wrote an extensive review on the exciting new field we call “precision...approach: Phenotypic Characterization of a Comprehensive Set of MAPK1/ERK2 Missense Mutants. Brenan L...

...And there are the usual transfection drawbacks: transient expression and needing to work with a transfectable...For cloning, Zetsche et al used four oligos that consist of direct repeats and crRNA.  Similar to a jigsaw...

...monoclonal Cas9 background because you’ll have more consistent levels of Cas9 expression between experiments...pAAV-hSyn-DIO-hM4D(Gi)-mCherry Our current AAV inventory consists almost entirely of viruses expressing DREADDs ...

... effects. However, these methods can be labor intensive and quite time consuming.   As Cas9/gRNA activity...several systems to do this but most are labor-intensive, require multiple fluorescent proteins, and aren...

...Base editors make specific DNA base changes and consist of a catalytically impaired Cas protein (dCas or... to base editor activity and editing speed and consists of the following components: A plasmid in the...

...inefficiently that no industrial operation would ever consider them worth the trouble.  Enter ELSA: E. coli Learning...for research.  We’ve also involved ourselves in extensive outreach programs to encourage and embolden the...

...is an extremely useful technique that provides insight into protein:DNA interactions. ChIP works by using...exploring these interactions, and can give valuable insight into DNA expression and protein interactions, including...

... ultrasound contrast. At a very low volumetric density and with a resolution of less than 100 µm, the ...probable mature human miRNAs. Weiss’s collection consists of a library of reporter plasmids, each containing...

...the displaced strand.   Figure 1: EvolvR consists of an error-prone polymerase (PolI) that is fused...provide a better understanding of the key residues responsible for this protein-drug interaction.   The Dueber...

...selected to participate on this panel, but my consistent writing for the Addgene blog (~1-2 blog posts...social media, but how you use social media to consistently engage with others about your topic(s) of interest...

...a nanobody.  AsLOV2 is a light-oxygen-voltage sensing domain from the oat plant (Avena sativa). In darkness... of more OptoNBs would allow for more uses both inside and outside cells. References Gil, Agnieszka ...

... and gotten unusual or unexpected results? Or considered why DpnI will degrade your template DNA from ...level, the potential for methylation should be considered when digesting DNA. Why? Well, even though Dam...

... in the human genome. The pair has focused considerable attention on transcription factors, starting... Myers: Only a small percentage of the proteins inside a cell have a commercial antibody against them....

...phosphorylated MEK1/2 levels, suggesting PP1/PP2A is responsible for this MEK1/2 negative feedback. 3. Studying...Silvia Carrasco, and Markus W. Covert. "High-Sensitivity Measurements of Multiple Kinase Activities in...

...be used to measure your fluorescent protein’s propensity to aggregate. The basics and pitfalls of these... the best way to test a fluorescent protein’s propensity to homodimerize. Several studies have used the...

...toxin-antitoxin system encoded by the ccd operon, which is responsible for plasmid maintenance during cell division....E. coli do not contain the F plasmid (and are considered F-); however, there are a few popular lab strains...

...even come [to grad school]!” He asked if she had considered other careers, which she hadn’t.  He then said.... “I was lucky to have been able to make that transition really smoothly,” says Susan. After being in ...

...increase activity and decrease the gRNA size by intensively remodelling the gRNA scaffold through five rounds...tools, the desired edits are templated by the 3′ extension of the pegRNAs. PEDAR is capable of correcting...

... transport miRNA from drug-resistant cells to sensitive ones. Though this process is not fully understood...malignancy, concluding that these cells may be considered "natural tumor suppressors." A growing interest...

...your network. If someone has connected with me, I consider them fair game for connecting to others.  Pro ...including and especially the hiring manager. This is considered intrusive and it’s still connecting with a stranger...

...Streak on them an E. coli  strain known to be sensitive to that antibiotic, and if the strain is growing...positive clone after these steps, you might want to reconsider your approach, and try a different cloning strategy...

...naturally in camels, consist of a single heavy chain, and they are small and stable inside cells. Nanobodies...

...abundance before and after a treatment. The library consists of a pool of lentiviral vectors with roughly 73...populations, they found that similar barcodes consistently made up the majority of the resistant populations...

...abundance of your targets isn’t too different, consider antibody affinity: stronger binding antibodies...membranes recommend re-wetting with methanol before rinsing with wash buffer, as dry PVDF won’t evenly re-absorb...

...the ‘quality’ of the gRNA sequence, including propensity for off-targets, secondary structures, etc., ...more pan-genome PAM flexibility, you’ll want to consider one of several Cas9s that recognize NG PAMs. The...

... crossing the blood-brain barrier.  Another consideration when picking ligands for chemogenetic studies...response you aim to manipulate. It is important to consider that the same receptor can elicit different responses...

... 1000000097) The Winslow lab barcoding library consists of 96 retroviral plasmids (based on the Murine...mammalian cells. The MuLE system kit from the Frew lab consists of 91 building block Entry vectors and 5 lentiviral...

...because it is non-pathogenic, well-studied and inexpensive. In addition, a complete set of well-annotated...different due to a whole genome duplication and extensive rearrangements in the S. cerevisiae lineage (7...

...scientists to confidently choose reliable and consistent antibodies for specific markers. The Screening...production methods    NeuroMab and others have an extensive library of monoclonal antibodies; Addgene collaborates...

... toolkit from Benjamin Field’s lab. Special Considerations There are a few special exceptions to the usual...constructs or certain cell environments). Also consider that a protein fusion may disrupt functionality...

...   Let me leave you with a few practical considerations: Design your gRNA to overlap the CpG of interest...interest. Having the CpG directly inside the gRNA sequence is a great guarantee that DNMT1 will be blocked...

...Polyclonal antibodies are produced in animals and consist of a mixture of antibodies recognizing many different...hybridomas.  First, the long-term stability, consistency between batches, and molecular definition of ...

... example, the CAG promoter, a hybrid promoter consisting of a CMV enhancer, chicken beta actin promoter...any template DNA and primers.  After performing extensive QC on that sample, we’ll then load it on to our...

...Special breeding cases Whether your mouse colony consists of genetically engineered mice with a single mutation...strains that have been crossed, there are many considerations for effectively breeding your mice. Because...

...institutions. The conversations often move quickly into sensitive topics and uncomfortable places, from sticky ethical...rapidly changing landscape of genetics and our consistent stream of events mean that there is never a dull...

...and mice with Nanoblades CRISPR has also been extensively used to create transgenic mice by microinjecting...post Resources on Addgene.org Check out our comprehensive CRISPR guide Find CRISPR plasmids and resources...

...lack all viral genes, including those that are responsible for integration into host chromosomes, further...to other gene delivery tools that are under consideration, recent advancements in various AAV technologies...

...in different fields that can provide different insight into an area - not just working within the established...collaborators can also help you get some critical insight into what you're doing wrong. All this helps to...

...using bacterial phage vectors with CRISPR/cas9 sensitizing gene cassettes. Thse vectors are engineered to...genes in the bacteria thus making the bacteria sensitive to known antibotics. These might be used on hospital...

...strengths and work effectively. Leadership responsibilities Being a leader in an organization, you need... different hats and came to understand the responsibilities that come with leadership. The NPA was a great...

... a company. Revolution Bioengineering is two responsible adults in their 30s who have quit their academic...below. We’re happy to answer questions about this transition (so far) and what we’ve learned.  Thank you ...

... increase efficiency of these systems, though consistent expression can still be a challenge. A multitasking...powerful than it is. The nuclease domain of Cas3 is considered weak, especially compared to the power of Cas9...

...which way your population needs to be shifted, consider your perpendicular lines. In this case, the population...applying very large compensation values (100+), consider what two markers your fluorescent markers are ...

...epitope, a phenomenon known as sequence context sensitivity (Schüchner, et al. 2020). In their study, some... newer Myc-tag antibodies appeared to be less sensitive to sequence context.    So, do Schüchner, et al...

...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...