... Antibodies Institute for Protein Innovation Antibody Collection Institute for Protein Innovation...Tag Antibodies Antibody Plasmids Welcome to the Institute for Protein Innovation (IPI) Collection! Here,...

...serotype testing: AAV1, AAV2, AAV5, AAV8 and AAV9... Viral Vector Packaging Service AAV Serotype Testing AAV ...AAV Viral Vector Packaging Service: Serotype Testing AAV These AAV encode fluorescent reporters and can ...Inventory . The catalog ID for the 20 µL sized testing sample is denoted with a .T suffix (for example... (for example, AAV1). AAV Vectors for Serotype Testing pAAV-CAG-GFP (Plasmid #37825) Description : Ready-to-use...

...CRISPR FAQs Zhang Lab's CRISPR Frequently Asked Questions You may also like... Zhang Lab CRISPR plasmids...expression, though it is possible for the plasmid to still express without the G. Because the G is only one...selection tool that should be helpful in minimizing or testing off-target DSB. Is it possible to target a single...the efficiency will be lower, although this might still be okay if you are introducing a selection marker...(gRNA) in pX330 / pX335 with BbsI/AgeI double digestion. As a successful insertion will destroy the BbsI... My genomic PCR didn’t work, do you have any suggestions? The Zhang lab recommends using the Epicentre...Cong et al. , Science 2013 ) do you have any suggestions? To date, I have never used the Ex Taq. In our...

...Depositor Collections Allen Institute for Cell Science Plasmid Collection Allen Institute for Cell Science Plasmid...The Allen Institute for Cell Science has deposited plasmids to generate fluorescently tagged proteins...Subcellular Localization Plasmids Protocols The Allen Institute for Cell Science has produced the first publicly...additional cell line information from the Allen Institute for Cell Science data portal (Link opens in a ...cellular structures. Addgene ID Plasmid Allen Institute ID Tag Protein Structure 87420 PXN-EGFP AICSDP...IPSC cell lines can be ordered from the Allen Institute for Cell Science Cell Catalog (Link opens in a... a new window) . Protocols The Allen Institute for Cell Science has several protocol videos available ...

...repository. Browse the University of Florida serotype testing panel for the eye and brain....Packaging Service AAV University of Florida Serotype Testing Panel for the Eye and Brain Viral Vector Packaging...Packaging Service: University of Florida Serotype Testing Panel for the Eye and Brain You may also like: Viral...by ddPCR, in vitro and (when possible) in vivo testing, and full sequencing of the final viral vector ... of glaucoma and their reversed cell entry characteristics. Gene Ther . 2020 Apr;27(3-4):127-142. PMID... AAV44.9-Based Vectors Display Exceptional Characteristics for Retinal Gene Therapy. Mol Ther . Jun 3;...

... Protocols Diagnostic Restriction Digest Diagnostic Restriction Digest You may also like... Restriction...unique but distinct patterns so that you get double confirmation. In the example above, digestion with either...Protocol for how to perform a diagnostic digest to verify plasmid size, verify insert orientation, and...specific piece of DNA from one plasmid to another. A diagnostic restriction enzyme digest takes advantage of ...quickly used to verify your plasmid. The goal of a diagnostic digest is to cut your plasmid into specific sized...all of an insert from it. Before beginning your diagnostic digest, you will need to select a restriction...Insert and Backbone Size The simplest form of diagnostic digest is one in which you just want to verify...

... you. Statistics – The basics Statistical analysis is what determines if there is statistical significance... several core factors of statistics relevant for many downstream statistical tests.   Independent variables.... 2). Assumptions – characteristics of a data set; the ‘rules’ a statistical test assumes your data...are statistically different. They are classic parametric tests and one of the simplest statistical tests...if the dependent variables worked synergistically, antagonistically, were additive, etc.). ANCOVA and....   Fig. 3: Question flowchart for choosing an ANOVA/MANOVA statistical test   Post hoc tests...described above. Violated assumptions and statistics Statistical power, the ability to correctly identify...

...we introduce you to the Journal of Emerging Investigators, an open-access journal that enables high school... the folks behind the journal and hear from a fantastic team of high school students who recently submitted... the interview with the Journal of Emerging Investigators Podcast breakdown Intro Chapter 1 - Introduction...Olivia Ho-Shing, and the Journal of Emerging Investigators. Chapter 2 - Getting high school students and...

... interest in “stingy science”.  Practicing thrift and conservation in the lab One stingy science project...me that most Addgenies make an effort to avoid plastic bags and to conserve resources where possible.  ....  At the lunch table, for instance, we have a plastic container problem…reusable containers pile up by...sink after lunch as fewer and fewer people use plastic bags.  We also try to recycle our discards according...about conserving resources in the labs. Disposable plastic materials are fairly inexpensive in this country... funded and we didn’t feel the need to reuse plasticware or think about conserving reagents and buffers...competent cells to save time and money. My other stingy science project started when I realized I was pouring...

...What are Acoustic Reporter Genes (ARGs)? Fluorescent and bioluminescent reporter genes are widely used...members of our lab - the Shapiro Lab - developed Acoustic Reporter Genes (ARGs), which are sets of genes...ultrasound nonlinearly, making them difficult to distinguish from background tissue unless destructive ultrasound...39006 produced the highest levels of nonlinear acoustic contrast in E. coli. We deleted an unannotated...the first-generation bARGs, poor expression and acoustic properties of these gas vesicles made non-destructive...expression, a smaller genetic footprint, and nonlinear acoustic scattering. The best candidate for this was the...the transfection mixture in HEK293T cells gave drastic improvement in expression. Stabilizing the gvpA...

...this signal is coming from.   Acoustic reporter genes (ARGs) Acoustic Reporter Genes (ARGs) are engineered...optimized for expression in E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference...ARG’s distinct ultrasound fingerprint can be detected. Bourdeau et al calls this method acoustic collapse...for serial acoustic collapse to aid in the development of ARG constructs with new acoustic phenotypes....but each of these reporters has its drawbacks. Acoustic reporter genes (ARGs) overcome these limitations...ultrasound. Challenges of in vivo imaging with existing bacterial reporters Limited Depth of Signal/Imaging... organs located deep in the body, like the gastrointestinal tract. Require Administration of Substrate...

...answer this question: Look at master's-level job postings  Look at master’s-level job postings on a job ...know if you should earn a master? Here are a few questions to help you decide if a master’s might be for ...website like Indeed. Do the job descriptions sound interesting? Is the work something you can see yourself enjoying... enjoying?  To find master’s-level postings, try adding “master’s” to your favorite search term, i.e. ...technical writer,” among others. Don’t discount job postings looking for PhD applicants since many will also..., pharmaceuticals, etc.) Do their jobs sound interesting? Is their career path one you could see yourself...master’s is a big decision, but hopefully the questions in this blog post can help you start to figure...

...science, but also science writing.  “And since I was still in grad school, I had the opportunity to take a ...science writing internship with the National Eye Institute (NEI), where she wrote about NEI research for ...skill set wasn’t matching up with the local job postings. “There were jobs that I didn’t seem to have the...Twitter feeds and dealing with social messages and postings,” says Susan. There are also team meetings and... Susan personally found this easier to do while still in school since opportunities, such as writing for...Addgene Other Resources The National Cancer Institute (NCI) has an internship similar to Susan’s NEI...

... et al., Neuron. 2019. PubMed PMID: 30704911. Acoustic reporter genes for noninvasive imaging of microbes...Article contributed by Lukas Morgan  Listen to the acoustic reporter gene podcast segment In recent years,...solution, Mikhail Shapiro’s lab has developed four acoustic reporter gene (ARG) plasmids to better monitor...the ARG plasmids at Addgene. For more details on acoustic reporters, check out this blog post. Bourdeau ... Cas9-expressing mice, and used the cells to reconstitute the immune system of wild-type mice. This resulted...

...fluorophores to distinguish between cells. Most flow experiments will use one marker to distinguish between live... Minibodies are engineered antibody fragments consisting of two scFvs bound to a CH3 domain. Fab fragments...measurable light or color in response to a specific stimulus, can be conjugated to antibodies in order to allow...machine or by X-ray film. HRP cannot be used to distinguish between different antibodies in the same reaction...the number of signaling molecules that can be distinguished in your application. The indirect detection ...molecule to epitope ratio. Secondary antibodies can distinguish between different species of origin, isotypes...between DNA and protein. Micrococcal nuclease digestion is used to shear the DNA in these assays. This...

...the other half to CIB1, light-stimulated heterodimerization can reconstitute the protein. This principle...examples of relevant variants. This list is not exhaustive - please browse our curated list of opsins here...neurons upon exposure to light through direct stimulation of ion channels. Naturally occurring channelrhodopsins... nonspecific cation influx into the cell when stimulated. Channelrhodopsin-2 (ChR2), the first widely ...domains from two different ChRs. These ChR variants still function as light-gated, cation channels resulting...Chronos High-speed, light-sensitive channel from Stigeoclonium helveticum 500 Chrimson, ChrimsonR Red-light...population, or activate different neural populations at distinct times. Temporal considerations. Temporal precision...

...oligonucleotide stitching, in which pieces of DNA that share no end homology can still be fused together...Gibson Type IIS Ligation Independent Cloning Oligo Stitching Molecular cloning or the creation of recombinant...containing precise 5' or 3' single-strand overhangs (sticky ends), or no overhang (blunt ends). Two pieces ...is done via a single recombination reaction, drastically simplifying the process and reducing the amount...rapidly shuttled into any compatible Gateway® Destination vector, which contain attR sites via LR clonase...clonase enzymes. Thousands of Gateway® destination plasmids have been made with different promoters, tags...are two-fold. First, the entire cloning step (digestion and ligation) can be carried out in one reaction...

...PMID: 25398340 Walton, R. T., Christie, K. A., Whittaker, M. N., & Kleinstiver, B. P. (2020). Unconstrained...Publications CRISPR Overview Bacteria have an interesting adaptive immune response that detects and eliminates...DNA with no homology elsewhere in the genome. Realistically, most gRNA targeting sequences will have additional..., a large portion of the Cas9-induced DSBs will still be repaired via NHEJ. The resulting population of...integrase landing site ( attB ) and the other to stimulate DNA repair. This can result in more efficient ...activity. Knockout libraries often target 5′ constitutively expressed exons, while activation and repression...repression, many CRISPR epigenetic tools require constitutive expression of the dCas9 fusion and/or expression...

...Description CMV Constitutive Strong mammalian promoter from human cytomegalovirus EF1a Constituitve Strong mammalian... factor 1 alpha CAG Constitutive Strong hybrid mammalian promoter PGK Constitutive Mammalian promoter ...Description T7 Constitutive but requires T7 RNA polymerase Promoter from T7 bacteriophage Sp6 Constitutive but requires...Inducible Tetracycline response element promoter U6 Constitutive Human U6 nuclear promoter for small RNA expression... simpler than eukaryotic transcription bacteria still have complex systems of gene regulation, like operons...polymerase Promoter from Sp6 bacteriophage lac Constitutive in the absense of lac repressor (lacI or lacIq...