...this situation: You’ve spent hours scouring a company’s LinkedIn pages, given yourself the bathroom mirror...socks, persevered through a nerve wracking commute complete with a disabled train, and now you’re sitting ...the candidate has what it takes to thrive at the company and add value, not just “do the job”. When they...these responsibilities. If you are in charge of completing many meticulous protocols or tasks, you’ve probably...

...gross comparison of AAV.PHP.eB and AAV.PHP.S to their parental vectors. To quantify this comparison, capsids....PHP.S had 1.5- to 2-fold higher GFP intensity compared to the strains they were derived from. They also...research. Visit Our AAV Guide Page for info on AAV components, common uses of AAV, and much more! Find ready-to-use...

...his program. Initially, Provost made his labs completely open-ended, allowing every student to work on...Kowalski et al., 2016; Scott, 2021). In fact, completion of Provost’s CURE increased students’ confidence...them to near homogeneity using a 6XHis tag, and compared their specific activities by measuring NADH production...Poirier, V., Pellettieri, J. 2020. The exon junction complex is required for stem and progenitor cell maintenance...

...lentiviral vectors! Lentiviral vector history and components In the early 1990s, researchers developed viral...system, the possibility of creating a replication-competent virus via recombination between the transfer and...recombination events necessary to create a replication-competent virus. Second, Tat is eliminated through the addition...Lentiviral barcoding libraries have been used for complex applications like lineage tracing in heterogeneous...

...restriction enzyme digest; however colony PCR can accomplish the same thing in less time and for less money... few ways you can do this. If you are going to complete your colony PCR analysis in the same day, you ... product size on a gel   Now that your PCR is complete, it’s time to run the products on an agarose gel... larger sized products for positive clones (+) compared to negative clones. Finally, orientation-specific...

... an RNA editor The Zhang lab envisioned a two-component RNA editor: a Cas13 enzyme fused to an RNA adenosine...off-target editing. In one direct transcriptome-wide comparison, REPAIRv2 reduced off-target sites from the 18,385...edits by ~45% and off-target A-to-I edies by ~94% compared to RESCUE.   Addressing the potential use of RESCUE... Abudayyeh, Omar O., et al. "C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector...

...orders were affected by the COVID-19 pandemic, we compared the number of plasmid orders and new COVID-19 ...the likelihood in our Poisson process into two components: 𝜃x(t) and 𝜓z(t). 𝜃x(t) describes the exponential...effects fairly well. However, we observed one complication in the China data – there is a period of 1 month... lockdown in China, then the observed data is compatible with an infinity-fold reduction in activity due...

...real time. When the PCR is complete, the results are analyzed by comparing the threshold fluorescence ...amplification reaction to take place.  When the PCR is complete, a droplet reader extracts the droplets from the...Absolute determination of single-stranded and self-complementary adeno-associated viral vector genome titers ...

...is ideal for tracking large collections. Many companies, such as Thermo Fisher and Micronic, provide 2D... time. 1D barcodes can be purchased from many companies such as Corning. Tubes can even have specific ...specific barcodes etched or printed onto them by companies such as Tubewriter. Any lab can benefit from using...tubes themselves. Although we’ve developed our own computer system for tracking samples, it can be done easily...

...enChIP system enables us to express the CRISPR complex much easier in various cells, including primary...give to post-docs and graduate students as they complete their appointments in your lab?  Fujii: These ...survive as a researcher because of the tough competition for academic positions and poor grant situation...develop new technologies, hopefully they'll be completely different from what we are now doing in my lab...

...on OWW, including those for preparing your own competent cells. If you’re familiar with the process of ...service free of charge to scientists by allowing companies to pay publish their catalogs on the site. Like...of checking off each step as you painstakingly complete your experiment – one small victory in a long ...manage scientific data: Electronic lab notebooks compared with Google Docs and wikis - by Rory Macneil on...

...interior target sites. In a second step, the two compatible target sites are cleaved out, leaving behind ...a vector with the inverted DNA flanked by non-compatible target sites (Schnütgen et al. 2003). Flex vectors...within neuronal networks in living animals. To accomplish such cell specific manipulations in vivo, FLEx...knockout. In this experiment it is possible to compare neuronal morphology and synaptic connectivity before...

...can be difficult to construct and deliver when compared to gRNAs. Additionaly, cDNAs cannot be used to...Housden BE, Perrimon N, Collins JJ, Church G (2016) Comparison of Cas9 activators in multiple species. Nat Methods...transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517:583–588 . https://doi.org/10.1038/nature14136...Kadoch C (2018) Interrogation of Mammalian Protein Complex Structure, Function, and Membership Using Genome-Scale...

...Host range depends on whether the host has the components to recognize the ori and allow for replication...organisms. pBBR1-based plasmids and plasmids in incompatibility groups N, P, Q, and W are known to have replicons...list of the bacterial species these plasmids are compatible with.  Table 1: Non-exhaustive list of common... and Srivastava 2013. Family Replicon Compatible with Shown to work in IncN pCU1 Gram-negative...

...meets two conditions:  1) The sequence is unique compared to the rest of the genome.  2) The target is present...found. If you’re lucky enough to find a SpCas9 compatible PAM at your site of interest, that is the most...expanding number of Cas9s with looser PAM requirements compared to SpCas9. xCas9 has a PAM requirement of NG, ... proteins. The Synergistic Activation Mediator complex, generally the strongest activator, consists of...

... and the complexity of effectively deploying them in the lab somewhat mirrors the complexity of understanding.... Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult both to..., and jGCaMP8 viral preps distributed per year compared to other AAVs preps from 2016–2023. B) Inset showing...

...virus as self-complementary AAV (scAAV). However, scAAV vectors have limited size compared to single-strand...-multi (Walton et al. 2024). This platform is compatible with enrichment, single-cell sequencing, and ...virus’s most abundant proteins and a critical component for viral genome packaging. Addgene recommends...

...reaction creates an expression clone with all of the components necessary for gene expression.     The LR... are performed, the next step is to transform competent E. coli cells and select the positive clones. ...functional. Advantages of the Gateway cloning method Compatibility and flexibility Once you generate the entry ...have generated and deposited their own Gateway-compatible plasmids with Addgene. These can be used to express...

...target provided that (1) the sequence is unique compared to the rest of the genome and (2) the sequence...for the canonical S. pyogenes PAM sequence NGG compared to wild-type SpCas9, which also displays some ...CjCas9) is even smaller than SaCas9 and is also compatible with AAV delivery. NmCas9, another small ortholog...(2018) Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Nature 556:57–63 ....

...fluorescent tools for labeling structures and compartments in mammalian cells By Angela Abitua Genetically...are often used to label  specific structures, compartments, or specific localization of biomolecules within...usually much easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed...plasmids were expressed in SHuffle T7 Express competent E.coli (NEB) using IPTG induction. To biotinylate...