... CAG Calcium sensor, Cre-dependent GCaMP Kim , GENIE 104487 pGP-AAV-syn-jGCaMP7s-WPRE Syn Calcium sensor...pGP-AAV-syn-jGCaMP7f-WPRE Syn Calcium sensor GCaMP Kim , GENIE 135632 pAAV-S5E2-GCaMP6f E2 regulatory element Calcium sensor ...pAAV.Syn.NES-jRGECO1a.WPRE.SV40 Syn Calcium sensor RGECO Kim , GENIE 162375 pGP-AAV-syn-jGCaMP8m-WPRE Syn Calcium sensor GCaMP...Viviana Gradinaru's lab at Caltech. Viral... Viral Vector Packaging Service AAV Caltech Systemic Capsids Viral Vector...Vector Packaging Service: Caltech Systemic Capsids PHP.eB PHP.S PHP.V1 MaCPNS1 MaCPNS2 CAP-B10 CAP-B22... from the Viviana Gradinaru laboratory and the Caltech CLOVER Center (Link opens in a new window) . As...

...peptides with known subcellular localization patterns. Useful for co-localization studies....mEmerald-Paxillin-22 Focal Adhesions paxillin mEmerald* Michael Davidson 15233 Paxillin-pEGFP Focal Adhesions paxillin... RFP-zyxin Focal Adhesions Zyxin RFP Anna Huttenlocher 11908 pEGFP-N1 alpha-actinin 1 Focal Adhesions ...pPaxillin-PSmOrange Focal Adhesions paxillin PSmOrange Vladislav Verkhusha 79999 pZyxin-miRFP703 Focal Adhesions...Dorus Gadella 158004 pCMV-mGold-Calnexin-N-14 Endoplasmic Reticulum Calnexin mGold Francois St-Pierre 158002...Organelle Markers Fluorescent Proteins: Subcellular Localization More Fluorescent Protein Resources: Empty Backbones...Secretory Pathway Background The subcellular localization of a protein is often tied to its function, ...

...I’ve answered hundreds of phone calls and thousands of customer service emails in my six years as a senior...spend far less time answering help emails and phone calls, I feel compelled to share my years of accumulated...customer, can get the most out of your email or phone call. Though my experience is based solely on my time...texting, actually writing an e-mail or making a phone call is a big commitment when you’re busy. Your time ... A lot of companies and organizations selling biological reagents have a ton of data on the catalog page...it’s own tongue-in-cheek website.  If you are skeptical about using Google for serious science questions...thread or remove a good 5 minutes from our phone call. If your concern is based on data you have produced...

...source of information on recent advances in the biological sciences and techniques that simplify and expedite... help hone your blog post for our audience of biological scientists from a variety of different fields...

... use a mechanical method before your enzyme lysis. For cell cultures, enzyme lysis is typically sufficient...for your experiment. But if you understand the technical aspects of a western blot, you'll be able to understand... so let's dive in!      Figure 1: The technical decisions to make when designing a western blot...multiplexing is challenging and limited. Direct westerns typically rely on primary antibodies conjugated to fluorescent... antibody or loading sample, which may not be practical. Conjugated primary antibodies can be more difficult...lengthy protocol, require the use of hazardous chemicals, and are not always consistent. Precast gels, ...unless you buy specialty nitrocellulose membranes specifically developed to allow stripping. And as a safety...

...Chavez, researchers at the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 is an enormously...reassemble when exposed to specific UV wavelengths or chemicals (Polstein and Gersbach, 2015; Zetsche et al., ...much cheaper to use Cas9 activators. This larger scale of activation allows for exploratory-based research...pathways and genetic interactions. Additionally, large scale CRISPR-based screens can be used to gain insight... Tuttle and Alex Chavez! Marcelle Tuttle is a medical student at Tufts University and former researcher...researcher at the Wyss Institute for Biologically Inspired Engineering.     Alejandro (Alex) Chavez is a member...MC, Qi LS, Kampmann M, Weissman JS (2014) Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation...

...values – numerical risk allowance in a statistical test. Distribution - spread of data; typically, normal...Statistics – The basics Statistical analysis is what determines if there is statistical significance among ...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...analysis can provide information on specifically which groups are statistically different within your ANOVA/ANCOVA... data in ascending numerical value relative to quantiles derived from a theoretical distribution. If your... non-parametric statistical tests! Adding it all up  There are many, many statistical tests beyond the...

...newly developed calcium sensors. Selecting a calcium sensor Genetically encoded calcium indicators (GECIs...biosensors. Nature chemical biology. 2021 May;17(5):509-18. Inoue M. Genetically encoded calcium indicators to...Iino M. Genetically encoded fluorescent indicators for organellar calcium imaging. Biophysical journal...fluorescent proteins linked to calcium binding proteins (e.g., calmodulin or troponin C). Their fluorescent... monitor calcium concentrations and transients in living cells and organisms. Unlike chemical indicators...unusually fast calcium dynamics, whereas non-neuronal systems tend to have slower, but larger, calcium changes...sensitivity parameters. Sensitivity and Localization The calcium concentrations in different subcellular...

...names for technical support roles. Examples are Technical Service Representative, Technical Support Specialist...these departments from technical support. Technical support careers in brief Technical support careers are...troubleshoot problems at the bench. Specifically, roles in technical support make excellent use of the ...worked as a technical support specialist for 4 years. Read on for insights from my time in technical support...products successfully. In technical support you may have to answer technical enquiries such as: “Can I... areas of expertise in technical support positions. When applying for technical support positions, you...familiarize yourself with technical support. Even having personal experience with technical support through the...

... the types of viruses used as biological tools.   Viruses as biological tools In nature, viruses infect... why viruses are such wonderful scientific and medical tools. As scientists, we can manipulate what this...effectively infect a wide range of cell types that are typically ‘difficult to transfect’, such as primary cells...with whole organisms. Finally, viral delivery is typically straightforward across applications, while electroporation... cells. They accomplish this through nuclear localization signals which can traffic their viral particles...Packaging Capacity (~4.5kb From basic science to clinical applications, viruses are true bench to bedside...

...background, which means that each mouse is genetically identical and all mice of the strain are homozygous...create a limited but genetically diverse gene pool. Outbred strains are strategically bred to maintain maximum...using the same nuclease may not result in genetically identical mice. Thus, scientists must genotype each...organism used to understand mammalian traits and genetically engineered mouse models provide researchers with...useful and adaptable tools to perform basic and preclinical research. For scientists new to using mouse models...) National Institutes of Health (NIH) Swiss. Genetically engineered mice Mice can be engineered by inserting...into a blastocyst. There are several types of genetically engineered mice: transgenic mice, knockout mice...

...re working on a project where we aim to do large scale DNA preps with multiple samples without cross contamination...

...packaging, considering using this PEI Calculator. AAV packaging typically requires transfecting three plasmids... ratios right, you need to do a few calculations. This calculator can do the math for you and it’s simple... download the PEI calculator for AAV packaging Use the PEI transfection calculator for AAV packaging ...spreadsheet will calculate the following μg of each plasmid needed per dish or stack. Calculations are based...corresponding column in the calculator spreadsheet: We hope that this calculator will make it easier to plan...whole experiment And here’s the equation the calculator uses to give you the μl needed of each plasmid...experiments. Let us know what you think about the calculator in the comments section below. References 1....

...about Julie Cicalese  Ms. Cicalese served as the Chief Human Resources Officer for Harvard Medical School ...excited to welcome Julie Cicalese to the Addgene Board of Directors. Cicalese served as the Chief Human...Human Resource Officer for Harvard Medical School. The addition of Cicalese to the Board of Directors brings...When I was working in Human Resources at Harvard Medical School, Joanna was my colleague. She mentioned ... science but my prior HR experience at Harvard Medical School means I understand the needs of scientists...Resources Association, Association of American Medical Colleges and the Society for Human Resource Management...

...SPARC to image the calcium response in the dendrites of T5 neurons by placing the calcium indicator GCaMP6f...researchers to label just a fraction of a single genetically-defined subset of cells. By labeling fewer cells...While transgenic animals are commonly used to specifically manipulate a cell type of interest in an organism...Drosophila require heat-shocking the system or using chemical inducers of gene expression (del Valle Rodríguez...cells are not labeled. PhiC31, a recombinase. It specifically integrates an attP site into an attB site. In...intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, in the Drosophila optic lobe...GCaMP6f in the effector position, as well as to optogenetically control the depolarization of R2 neurons using...

...Here, we will explain how practical brightness is determined and why practical brightness is a better criterion...criterion than theoretical brightness for selecting the brightest fluorescent protein. Theoretical brightness...quantum yield (QY) respectively. The theoretical brightness is calculated by multiplying EC by QY (EC*QY, ...higher the number, the higher the theoretical brightness. The theoretical brightness often appears in tables...proteins based on their practical brightness in your experimental set up. The practical brightness takes all...same reference protein) under identical conditions, a ranking of the practical brightness can be made (Goedhart...which is a key determinant of the practical brightness. Therefore, practical brightness provides a better ...

...expected when we originally sent out the call for applications. Cervical cancer screening in Ghana Gladys is... she can use as positive controls in PCR-based cervical cancer screens. This third grant represents the...is currently working to enhance cervical cancer screening in Ghana. While pap smears and visual tests ...Papillomavirus (HPV). As Gladys wrote in her application, “Cervical Cancer is the leading cause of cancer related ...in other developing countries” and “Almost all cervical cancers are caused by persistent infection of ...controls To start her work, Gladys will improve cervical cancer screening for a limited group of Ghanaian...including PCR screening for HR HPV alongside the cytological tests currently in use. Gladys will use plasmids...

...Addgene can be found on our Contact page. You’ve called. You’ve emailed. And Addgene has been there to ...plasmid questions. But until now, you’ve always had to call the United States - Cambridge, MA to be exact. Today...collaboration with LGC Standards. And now you’ll be able to call us - Benoit Giquel and Emma Markham - the newest...the University of London and the University of California, specialising in Molecular Biology. She has worked...Genetics. Since then she has worked in the the UK, California, Africa, and Australia, viewing science as a ...   Europe, we are all geared up to answer your calls, emails, or maybe even visit your lab! We look forward...

...recent paper from Ed Callaway’s lab (Fischer, KB. 2019) set-out to systematically test the potential causes... was not present (Kuhlman, SK. 2008). While theoretically this strategy should work, these vectors suffered...expression.  As such, researchers developed new tools called FLEx/DIO vectors where the transgene is floxed ...conditions used to minimize recombination that are typically employed during viral plasmid amplification (Fischer... the researcher's system before beginning large scale experiments. This will help to ensure the accuracy...doi.org/10.1128/jvi.02213-17 Fischer KB, Collins HK, Callaway EM (2019) Sources of off-target expression from...IR (2020) Monosynaptic Tracing Success Depends Critically on Helper Virus Concentrations. Front Synaptic...

... you to do customized genetic modification on a scale that was really not possible before,” said the Broad... matter of a few days. Their library, which they call GeCKO, targets 18,080 human genes with 64,751 unique...aimed for 300 cells per construct, but in some biological systems one “might need larger numbers.” Further...Lander ES. Science. 2013 Dec 12. (PubMed). Genome-Scale CRISPR-Cas9 Knockout Screening in Human Cells. Shalem...