...studies have shown that the conditions Sotos-type 2 and Marshall Smith Syndrome are due to mutations in...accurate diagnosis of NFIX-related syndromes (Sotos-type 2 and Marshall-Smith) and could be expanded to NFIA-...lines and use these lines to study how immune processes and biological pathways are altered in PMM2-CDG...

... that all look exactly alike.      Figure 2: Reagent gradient. Here, the lanes are lightly marked...guide. The gel is then removed from the membrane (2) and the membrane is cut so each lane is its own piece...in the bands, you may want to consult Tables 1 and 2 in our Technical Design of a Western Blot post to ...the antibody’s dynamic range. Specifically, the 1:2, 1:1, and No dilution samples all have approximately...conditions are 4 °C overnight or room temperature for 1–2 hours, there is some evidence that antibodies may ...high background or low signal during the imaging process, try adjusting your exposure time first, since ...

...antibiotic selection. Essentially, this gene is a 2-in-1 providing both bacterial and eukaryotic selection...Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage/gei016I .... During the propagation and/or transformation process, some bacteria will either never receive a copy...antibiotics listed below affect continuous cellular processes (e.g., protein synthesis), while amp only affects...

...with neuron-specific expression of dCas9-VPR (Figure 2), a popular Cas9 activator developed by the Church...ubiquitous ones such as EF-1α and CAG. Figure 2: Dual lentivirus system with ubiquitous expression..., studying the genetic underpinnings of these processes has been complicated by the lack of precise genetic...undifferentiated cells at the end of the screening process. However, while the neuronal marker TUJ1 is useful...

...green squares, purple circles and blue triangles. 2) An antibody immobilized on a bead binds specifically...protein G, a novel IgG-binding reagent. J Immunol 133(2):969-974. Hjelm H, Hjelm K, Sjöquist J (1972) Protein...to immunoprecipitation and simply refers to the process of anchoring an antibody, often called the capture...protein interactions, and limit high throughput procedures. However, agarose beads are often a more cost-effective...

... reduce the background staining (Table 2).      Table 2: Secondary antibody selections for a four-antibody...have been cross-adsorbed. The cross-adsorption process filters out off-target antibodies increasing specificity...

...wheat germ, HeLa, and Leishmania (Gagoski, et al 2015[2]). The proteins encoded in this set range in size ...to generate Illumina sequencing libraries using a 2-step PCR-based protocol and are compatible with single-end...decrease false positives observed in the cloning process, making it easier to produce the constructs you...novel and powerful pyranopyridine derivatives." Proceedings of the National Academy of Sciences 113.13 (2016...-time observation of metabolite production." Proceedings of the National Academy of Sciences 113.9 (2016...pCri system to simplify the protein purification process by making it easy for a researcher to quickly generate...on the system Easing the Protein Purification Process with pCri Cell-free protein expression kit Cell-free...

...break), increasing editing efficiencies 2-3-fold. However, the process of double nicking increases indel formation... editing as efficient as possible. Figure 2: Decision tree for selecting a PE6 variant. PE = prime...using paired prime editing. Nature Biotechnology, 40(2), 218–226. https://doi.org/10.1038/s41587-021-01025...the template and primer binding site, increase processivity, or improve thermostability. What came out on...repair (MMR) complex, to temporarily inhibit this process and promote copying of the edited strand to the...to the numerous innovations that streamline the process of prime editing, the Liu lab contributed an improved...

...whether the results include knockout data.    Figure 2: Search options in the Antibody Data Hub. Data reports...antibody worked in your application. The submission process is easy, with simple, descriptive fields, drop-..., and can be saved at any point throughout the process, so you can work through it at your own pace. To...

...gene activation, with some genes experiencing around 2-fold activation levels. Although other methods have.... This method has been shown to activate genes at 2- to 3-fold higher than SAM, SunTag, and VPR (Zhou ...activation across multiple loci was an arduous process. When using zinc finger proteins or TALE proteins...

... digest and compare it to some reliable controls. 2. Viral stocks may be sensitive to freeze thaw cycles...PMID: 24003179. PubMed Central PMCID: PMC3760253. 2. Rohwer, Forest. "Global phage diversity." Cell 113.2...volume. Always remove packaging cell debris before proceeding to ultracentrifugation to avoid contaminating...

...estimate transformation efficiency (Figure 2).    Figure 2: Example of a serial dilution to estimate... reduce recombination during the amplification process. In liquid culture, all of the bacteria can grow...

... PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an insertion locus-specific gRNA The gRNA ...Find the plasmids from this publication at Addgene. 2. Dalvai, Mathieu, et al. (2015). “A Scalable Genome-Editing...MMEJ doesn’t get a lot of press. However, this process accounts for a percentage of the mutations seen...

...from GFP and the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising..., V163A EBFP F64L, S65T, Y66H, Y145F   Table 2: Functional role of specific mutations in GFP derivatives... enabling the study of dynamic, physiological processes. Re-engineering GFP to increase its range of color...

...Information Science and Technology 60.4 (2009): 858-862. 2. Van Noorden, Richard. "Interdisciplinary research...Librarianship and Scholarly Communication 2.2 (2014): 2. Additional Resources on the Addgene Blog 6 Tools...others for failing to find our work?  I study processes for diagnosing and resolving organizational performance...impact of their contribution. It offers a 5-step process and several examples for creating effective research...

...citizens enter into in order to protect their rights (2). Less formally, governments are the organizations...science education and literacy that this changes.  2) Lumping all scientists (and consequently their results...politics? Science is an apolitical process for seeking knowledge. The process begins with a testable hypothesis...hypotheses and the cycle begins again. No part of this process is inherently political so why is there a march...scientists as well as other citizens. Thus civilization proceeds in an informed fashion. For science to be useful...

...faster when using a dual guide system.   Fig. 2: Size screening for large deletion events.   ... Biol 17: https://doi.org/10.1186/s13059-016-1012-2 Find more CRISPR Resources at Addgene: Read our CRISPRGuide...looking for, you are ready to begin the screening process. Want to set yourself up for a screening success...then a single, bulk validation may be enough to proceed with your experimental plan. Validating knockout...

...Halloween is only 2 days away and here at Addgene we couldn’t be more excited! As a small, tight-knit...suggestions for this year’s teams that may be procrastinating choosing their costume?  As our organization...

...First, the light-responsive molecules of cryptochrome 2 are the chromophores flavin-adenine dinucleotide and...of optogenetic control of endogenous cellular processes." The LITE system can now be applied to unraveling...genetic and epigenetic players in normal biological processes and disease states. While LITE plasmids have clear...

...would label everything but the CD8+ T cells (Figure 2). Even better, we can use the same fluorophore for...gating on the negative population.     Figure 2: Dump gating labels all unwanted cell lineages with... for entry of the antibodies. Here, a two-step process is employed: the cells are first fixed and then...phosphatases, and ubiquitin ligases; and many more. This process, however, leaves the nucleus intact. If markers...not need to use a separate fix/perm cytosolic procedure in that case.     Figure 3: For intracellular...