...control, including AAV titering by probe-based droplet digital PCR, in vitro and (when possible) in vivo...review performance data of each of these serotypes, please refer to Addgene’s Data Hub and the Caltech CLOVER... serotype will work in your mouse line, see Supplementary Table 3 (Link opens in a new window) . Browse...using the Caltech serotypes in future publications, please acknowledge Viviana Gradinaru and cite as follows...would like to see available in viral prep format. Please note this does not guarantee viral vector packaging...

... 12 GC/mL for the AAV2 serotype, as titered by droplet digital PCR (ddPCR) . In some cases, we may expect... sequence verification, may not yet have been completed at the time of shipment. If any discrepancies ...for any additional customs fees or taxes, and to complete any additional customs documentation that might...for the import of biological research materials. Please be aware that additional customs charges may be...

...microscope, or detect when the protein is present in a sample--just to name a few. Traditional sources of antibodies...individual clone targeting a single antigen. An example of R-mAbs is the NeuroMab collection - monoclonal...single-domain antibodies (also known as nanobodies). For example, scientists often use plasmids encoding scFv fragments...recombinant antibody fragments in the form of scFvs A simple vector system to improve performance and utilisation...

... Figure 2: Examples of simultaneous multiplex IF labeling of rat brain samples from the James Trimmer...Figure 1: Schematic of multiplex immunofluorescence approaches. A) Sequential Multiplex IF, where one target...you will need to start with a fixed sample. Depending on the sample type, this might be frozen or formalin-fixed...al, 2022 for a sample protocol.)  Simultaneous IF Depending on your needs, your sample, and targets, you..., or you can make your own. For example, to label the rat brain sample shown in Figure 2A above, Andrews... get completely washed away and leave no signal. If you see lots of extra signal in these samples where...set of samples where you label only one target in each. Compare these individually-labeled samples with ...

...vector encoding the MS2-P65-HSF1 activator helper complex with a 2A Hygro resistance marker (EF1a-MS2-p65...Name Insert Antibiotic Resistance Purpose PI 17446 pLenti CMV GFP Hygro (656-4) GFP Hygromycin 3rd gen lentiviral...Suggestions Don’t see what you are looking for? Please let us know what you would like to see available... Suggestion Form (Link opens in a new window) . Please note this does not guarantee viral service, but...

...particle (a virion) are generally divided among multiple plasmids. Some plasmids (typically called packaging...lentiviral plasmids to express your CRISPR gRNA? Please see our CRISPR Empty gRNA plasmids table and search... See plasmid 17618 for GFP plasmid. Cheng 17452 pLenti CMV Puro DEST 3rd Gateway destination plasmid with... See article for more variations. Campeau 39481 pLenti-puro 3rd CMV driven expression of cDNA. Puro selection...

...for multiplex genome editing.  For a few reasons, Cpf1 is a simplified system for editing multiple targets...protospacer Multiplexing CRISPR-Cas9 options before Cpf1 crRNA array Prior to this new Cpf1 multiplexing method...allow for expression of multiple gRNAs (i.e. Csy4 cleavage sequence, tRNAs, multiple individual promoters...Cpf1 counterparts. Table 2. Cas9 Multiplexing Options  Multiplexing Method Delivery Method Advantages...Xie et al  Multiplexing crRNA expression with Cpf1  The key advantage of multiplexing crRNA expression... chart below for a summary of multiplexing efficiency in vivo. Multiplex gene editing with CRISPR-Cpf1... CRISPR technologies.  It’s a simple and effective method with multiple applications (in vitro, in vivo...

... cloning techniques with higher efficiency and simpler procedures have been developed. Some of these include... excessive time requirements. FastCloning is a simpler yet reliable cloning technique that was developed...insert with an additional 15-17 bases that are complementary to the overlapping vector ends. See an overview...an hour at 37°C. This should be enough time to completely digest the template DNA. Note—templates must ...

...end up with poor droplet quality. Using a droplet generator, the sample and the droplet generation oil ...take place.  When the PCR is complete, a droplet reader extracts the droplets from the plate and measures...reagents (and time)! AAV titering using droplet digital PCR Droplet digital PCR involves partitioning a PCR... approximately 20,000 droplets using water-oil emulsion technology. Each droplet contains the ingredients...emulsion containing approximately 20,000 droplets. Each droplet contains the material required for a mini...each one. Droplets that fluoresce contain the amplified target sequence. After all of the droplets have been...negative (gray) droplets. The no template control wells should have very few positive droplets. In the image...

...sites. Modifying multiple genes at once. Using multiplexed gRNAs can target multiple locations in the ...CRISPR makes it easy to target multiple loci - a concept called multiplexing. Since CRISPR is such a robust...you add multiple gRNAs on one plasmid. Sound good? Addgene has many tools to help you multiplex - we’ll...Drosophila, and zebrafish!   Why use multiplexed gRNAs? By expressing multiple gRNAs on the same plasmid, you...clone and transfect multiple promoter-gRNA constructs in order to target multiple sites! In this post,...we'll cover Cas9 multiplexing options, but also check out our blog post about multiplexing with Cpf1.   ...  Figure 1: Multiplexing allows researchers to express multiple gRNA from a single construct. DNA...

...scalability and efficiency of multiplex gene editing. Moreover, existing multiplex strategies have been facing...developed DAP arrays for efficient multiplex base editing (MBE) and multiplex prime editing (MPE) with only...Schematic of multiplex gene editing with DAP array   Use DAP arrays can enable efficient multiplex gene ...array for multiplex gene editing applications, we provide detailed manual and online design examples of using...natural CRISPR locus of a bacteria host encodes multiple guide RNAs (gRNAs) on a single array to target...leveraged such host-defense mechanisms to enable multiplex gene editing in a variety of cells and organisms...from a single array, has been a top choice for multiplex gene knockout or regulations, it lacks proper ...

...Fluorescent proteins (FPs) are also the foundation of multiple important technologies, including FRET and optogenetics...treatment can photoconvert GFP from green to RED! This simple technique has been shown to work in plant, Drosophila... like PsMOrange and mEos add another layer of complexity to imaging, as they allow one to monitor compartment...

...virus Mammalian constructs Due to the size and complexity of the collection, all available Ginkgo SARS-...information on ordering from a for-profit entity, please see our Plasmid Distribution to Industry page. ...top General Plasmid Tools SHERLOCK and DETECTR - Simple, portable, and inexpensive platforms to reliably...Collections of COVID-19 related articles spanning multiple publishers can be found here: ASM COVID-19 Research...

...dysregulation event. Diseases are often complex, with multiple layers of dysregulation from either genetic...to each cell, future applications could deliver multiple TFs to each cell, allowing for combinatorial overexpression...the specific dysregulation in the disease. For example, very few diseases manifest as a single mutation...

...author “Coping with Difficult People,” identified seven types of difficult people: Hostile-aggressives, who... are a few principles that will help you resolve most conflicts, even with difficult people. If I had ...supervisor, or do you have to work with difficult people? Each time I ask this question at my workshops,...had to use one word for to summarize these principles it would be “assertiveness.” 7 strategies to become...confidently while also considering the needs of other people. Some students and postdoctoral fellows are hesitant..., especially if their supervisors are difficult people. The following seven strategies will help you to...frequent sources of conflict is miscommunication. For example, you might misunderstand your supervisor’s suggestions...

...Dehydrogenase CUREs Community (MCC) Collection Pleiades Promoter Project: mini-promoters to drive selective...FreeGenes Project Open Enzyme Collection Screening Multiplexed Overexpression of Regulatory Factors (MORF) Collection...Factor Plasmids Terminator Plasmids Viral Vectors pLEG/pREG Modular Viral Vector System - Dankort...

...Cas enzymes that form a complex called Cascade ( C RISPR- as sociated c omplex for a ntiviral de fense...instead recruiting the transposase (comprised of multiple components) to target locations in the genome ...

...Cascade ( C RISPR- as sociated c omplex for a ntiviral de fense) complex, comprised of a combination of ...difficulties in getting all components of the Cascade complex to express equally. To combat this, some CRISPR-Cas3...

...design GGA overhang sets, particularly for high-complexity reactions? The researchers over at New England...data-driven approach might allow for an increase in the complexity. In 2018, they published a paper profiling four...2022).     Figure 1: Workflow for high complexity Golden Gate Assemblies. Image courtesy of New... three DAD tools used were key for ultra-high-complexity assembly. While the traditional rules of thumb...data-driven approach was needed to massively expand complexity. The three tools developed, outlined below, allow...traditional design rules) for successful high-complexity Golden Gate Assembly reactions. NEBridge Ligase...of overhangs, stat? GetSet can do that for you, complete with predicted fidelity. You can specify number...

... been used to date . If you have any questions, please write to us in the comments section.  FX cloning...important microalgae: intrinsic suicide switch implementation in cyanobacterium Synechocystis sp. PCC 6803...directly engages the ribosome for translational complex assembly. PhD student Dejan Gagoski used SITS ... research. If you’d ever like to deposit a kit, please contact us at [email protected]. ...