...tissues. This inversion creates a template that’s suitable for PCR amplification using pre-designed primers....increase transduction efficiency, none allow for simple and efficient transduction of both the CNS and ... not lost in a jumble of fluorescent neurons. A simple but flawed approach to multi-color labeling is ...

...Technique What It Locates ChIP-seq Proteins (for our example later, histones) ChiRP-seq RNA Hi-C DNA (genome-to-genome...appendix DNA is a 3D fractal). Enhancer biology is a complex and dynamic field. We’re going to focus on a tried...People are wrong. In reality, cellular DNA is a complex three-dimensional globule of chromatin. Chromatin...Bedtools getFasta. These regions and their reverse complements were parsed for spCas9 PAM sites (NGG) and then...

...is a terrible disease that is, unfortunately, completely preventable. Mainly caused by exposure to asbestos...that there are some genetic factors involved. For example, multiple studies show that certain mutations of...positive potential for treating mesothelioma. For example, patients who have peritoneal mesothelioma – that...immunotherapy that work in different ways. For example, a new drug known as pembrolizumab (Keytruda) causes...

...become resistant somehow overtime. One fantastic example of intrinsic resistance would be Mycoplasma species...Figure 1): Over-prescription of antibiotics (for example, giving antibiotics for an infection that isn’t...treatment regiment might wipe out bacteria that are completely susceptible to a drug while a full course of ...likely to develop over time. It isn’t feasible to completely eliminate the development of resistance; it’s...

...camera or tracking software. Locomotion is a highly complex process on the level of individual neurons and ...motivational and other aspects may be involved. For example, an animal moving around to explore the environment...the elevated plus maze, the light-dark box is a simple device comprised of a brightly lit (risky) and ...behavioral neuroscience to reveal insights into the complex relationship between genes, neurons, and behavior...

...the anticodon,  a three nucleotide sequence complementary to the mRNA sequence. The 3’ end of  the tRNA... the 20 amino acids and three stop codons. (B) Example of codon usage among different organisms. There...prokaryotic systems or yeast (Lanza et al., 2014). For example, it may be worth codon optimizing a human gene ...optimized for expression in mammalian cells. For example, Henry Lester’s lab optimized the C. elegans GluCl...

... list of specific structures within cells. For example, to improve nuclear localization of fluorescent...//doi.org/10.1101/160374 Nanobodies- there's a simple method for that! By Michelle Cronin The benefits...Kimiko Hara, and Kazuo Umemura have described a simple method which alleviates the inclusion body problem...problem and makes nanobody production in E.coli as simple as following a DNA miniprep protocol!  First, the...

...only occur through editing by a APOBEC-Cas9n-UGI complex. So how exactly does this work? Figure 1:...DNA editing efficiency of APOBEC-Cas9 editisome complexes in real-time (St. Martin et al., 2019). To create...DNA editing efficiency of APOBEC-Cas9 editisome complexes. The eGFP reporters rely on the correction of ... single base editing by APOBEC-Cas9 editosome complexes. Scientific Reports 9: . https://doi.org/10.1038...

...Schumacher, 2012). This concept is a little more complex than simply competing for replication control factors... daughter cell during cell division. They don’t simply rely on chance (or random diffusion) to make this...some types low copy plasmids during cell division simply does not allow for enough time for correct partitioning...

... 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...broadly useful vectors, a process that isn’t as simple as it seems. Take, for instance, the GCaMP-encoding...

...within the scientific community, as they allow for simplified transformation processes, and can be used by ... the others are functional in other hosts. For example, the pJD4 plasmid harbors distinct origins (ori1... load (Meyer 2009). Figure 1: Idealized examples of a narrow host range plasmid with only one ori... hosts in which proteins can be expressed. For example, scientists can incorporate interchangeable antibiotic...

...granularity, a proxy for nucleic complexity of the cell.  The sample will have been labeled using a panel...parallel.  Let’s imagine we’re looking for cells in a sample that are negative for marker A and positive for...Anderson, D. (2015). Neutrophil Responses to Sterile Implant Materials. PloS One, 10, e0137550. https://doi....

...Construction of solid matrix-antibody-antigen complexes containing simian immunodeficiency virus p27 using...structures of CRBNmidi alone (A) and in a ternary complex with the multiple myeloma drug mezigdomide and ...on its own, with drug ligands, and in ternary complexes with other proteins, establishing CRBNmidi as ...first use of CRISPR/Cas9 in M. abscessus. The simplicity and efficiency of this dual-plasmid approach ...

...targeting the genomic site of interest. The donor template is supplied separately. To use this system, you.... The optimized plasmid curing method was much simpler and took only ~32 hours instead of ~60 hours. ...strategy for plasmid curing, providing users with a complete editing process. The MUCICAT system While the ... pQCascade) with 4 rounds of transformation to complete the N-acetylglucosamine strain library construction...

...sequence of interest (red) is amplified using Recombinase Polymerase Amplification (RPA) or Reverse-transcription...RT-RPA), respectively. Amplified nucleotides are combined with Cas 13a in complex with crRNA targeting ...

...work in cell culture. The problems with cell lines Simply put, the majority of cell line problems come from...when it is important to check their identity. For example, perhaps a working cell bank has been made by a...monitor any changes. Your tracking system could be as simple as a piece of paper on the front of the incubator...passage number, cell line, and date, which you complete each time you passage the cells (as well as recording...

...often have absolute position requirements. For example, a KDEL ER retrieval sequence only functions at... even disrupt fusion protein localization. For example, the formation of inappropriate inter-chain disulfide...disulfide bonds between FPs blocks Golgi complex-localized fusions from properly exiting the ER and fluorescing... that can be used in various combinations. For example, with standard fluorescent microscope filter sets...

...proteins. lon protease: Strains where this is completely deleted (designated lon or Δlon) similary reduce...variety of sources, so fear not—there are a few simple troubleshooting measures that can help get you ...will not express in an IPTG induction strain for example, nor will a p15 plasmid be compatible with a pLys...better at 30°C or room-temperature, which is accomplished by growing your culture to the desired density...

...protein is cleaved by proteases. Nowadays, with ample structural information available, it’s clear that... better preserves the function of the POI. For example, the APT1-mVenus fusion localizes at the Golgi,... interact with other proteins or biomolecules. Examples of the successful insertion of a fluorescent protein...interactions) can be challenging. If a fusion can complement a knock-down or knock-out of the corresponding...

...scFv sequences have traditionally been cloned by amplifying the VLand VH antibody sequences of hybridomas...applications of scFvs: The SunTag system uses an scFv to amplify the fluorescence intensity of a tagged protein.....0000000000000145  Gąciarz A, Ruddock LW (2017) Complementarity determining regions and frameworks contribute... about the SunTag system which uses an scFv to amplify the fluorescence intensity of a tagged protein....