...donor and acceptor. FRET is often used to study (1) protein-protein interactions where each protein is...standard consisting of ECFP and EYFP connected by 1 flexible glycine- and serine-containing peptide linker...fluorescent protein. Plasmid Color Expression Description pPROEX Aqua Cyan Bacterial Expresses Aquamarine...

...affecting females at an incidence of approximately 1:10,000. It is characterized by apparently normal development...in transcriptional repression, activity dependent de-repression, chromatin remodeling, gene activation... the TRD contains a Nuclear receptor Co-Repressor 1/Silencing Mediator of Retinoic acid and Thyroid hormone...species (mouse or human) Mutation ID Plasmid Description Gene/Insert Mutations PI Return to top External...BrainSpan Atlas of the Developing Human Brain - includes developmental transcriptome information, prenatal...

...in mammalian cells or zebrafish). pMG10 (a '“2-in-1” ZFN cassette for expression of both a “+” and “-”...This page lists plasmid vectors designed to express nucleases in different cell types. Zinc finger arrays...Expression Vectors This page lists plasmid vectors designed to express nucleases in different cell types. ...zebrafish (can be used as a substitute for pMLM335 described in Foley et al., PLoS One 2009) pMLM292 (for expression...zebrafish (can be used as a substitute for pMLM336 described in Foley et al., PLoS One 2009) pMLM800 (for expression...Methods 2011 ) in mammalian cells or zebrafish as described by Moore et al. PLoS One 2012 and Cade et al.,...

...for any application TMN200 (200 mM sodium chloride, 1 mM magnesium chloride, 20 mM Tris at pH 8.0) Suitable...adeno-associated viral vector. Hum Gene Ther Methods . Feb;25(1):72-82. PMID: 24191859 Ye, et al. 2015. Safety and...name for ordering information. ID Name Promoter Description Category PI 157970 pTR-UF11 chimeric CMV/Chicken...

...Buenrostro JD, Regev A, Zhang F. Cell 2023 Jan 5; 186(1):209-229. PubMed (Link opens in a new window) . TF...identification during screening. Library ID PI Description MORF library 137000, 192821, 1000000218 Zhang...

...Expression mEGFP 488 507 34 6 Monomer (A206K) pmEGFP-1 - Mammalian Expression mEGFP-N1 - Mammalian Expression...TurboRFP-pBAD - Bacterial Expression tdTomato 554 581 95 4.7 1 hr Tandem-dimer pCSCMV:tdTomato - Mammalian Expression...-pBAD - Bacterial Expression mRFP1 584 607 13 4.5 1 hr Monomer pcDNA3-mRFP - Mammalian Expression pMXs-mRFP1... of Biomedical Optics, Feburary 2005, Vol. 10 No. 1, p. 14003 Chu et al. : Nature Methods, May 2014 Vol...Plasmids - Includes tagging with mCherry, mCitrine, mCerulean Davidson Lab Plasmids - Includes many N- and...fluorescence to obtain half-maximal value. Structure describes the monomeric or oligomeric state of the protein...System Plasmids Mammalian Golenbock Lab Plasmids - Includes tagging with CFP, YFP, and mRFP Tsoulfas Lab Lentiviral...

...researchers use ssDNA or dsDNA repair templates with 1. homology to the DNA flanking the DSB and 2. a specific...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Fully functional CRISPR...encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental considerations,...or search the tables below for CRISPR plasmids designed to introduce a DSB. Plasmids are available for...

...gene activation CRISPR/Cas9 Plasmid Variants Figure 1: Schematic of the CRISPR Collection. Plasmids 159654...can be designed using Benchling or using similar design tools and ordered as standard desalted oligonucleotides...phosphorylated oligonucleotides containing the guide target sequence: Up to two guides can be expressed...that utilize Golden Gate cloning to insert the desired guide RNA sequence by digestion of the backbone... #63798) Plasmids from Collection ID Plasmid Description PI Publications Kaufman, M. L., Goodson, N. B...

...Biology, 2(9), 537–541. DOI: 10.1021/sb400021j Figure 1: Multiplexing CRISPR knockdown to modulate expression...CRISPR guides. The first one binds to sfgfp with either 0, 10, 11, 14, or 20 matching nucleotides, the ...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts The David Bikard Lab at...mcherry with either 0, 11, 18, or 20 matching nucleotides. Together, these plasmids allow to explore the...

...research and innovation programme and EFPIA. Figure 1: Representation of RESOLUTE mission. The combination...proteomic interactions represent the framework that provides biological context. Assays monitoring impedance...sub-cloning into a wide range of Gateway-compatible pDEST vectors. ID Plasmid Gene/Insert Industry PI Pooled...solute carrier family of proteins. Library ID PI Description Human SLC Activation Library 132561 Superti-Furga...

... assembly reaction** 1 5 min., 37 ºC (2.5 μl) 687 100% 274,200 2,742,000 1 60 min., 37 ºC (2.5 μl)...expected complete sequence for the lacI/lacZ genes (1), while sequencing of white colonies showed a mixture...representative transformation plates obtained from 1-, 12- and 24-fragment assemblies of the lacI/lacZ ...lacZ cassette, and illustrates how the volume of the 1 ml outgrowth spread on each transformation plate can...fidelity studies conducted by Potapov et al. at NEB (1, 2). While T4 DNA Ligase, the mainstay of most biotechnological...over 90% for 24-fragment assemblies (Figure 3, Table 1). These improvements (5- to 12-fold increases in the... 16 ºC) x 30 (100 μl) 78 90.7% 783 9,792 Table 1. Yields and fidelities for Golden Gate Assemblies ...

...plasmids can be ordered via the links below: Table 1. dCas9 Activators sgRNA Dual Expression ID Plasmid...Table 4. Gateway Destination Vector ID Plasmid 48222 pAC90-pmax-DEST : Gateway Destination vector derived...the dCas9 protein provides the transactivation activity, the sgRNA component provides the specificity and...and can be transferred to Gateway destination vectors (e.g., pmax-DEST [48222] and (Link opens in a new...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts CRISPR-on is an RNA-guided... and is designed to target promoters or enhancers of genes of interest. Different flavors of activators...new window) Lifetechnologies Gateway Destination Vectors ) to adapt to different systems by LR clonase ...

...spike protein (MERS, CoV-1, CoV-2), modification, or variant. ID Plasmid Description Mutations Industry PI...expressing mCherry and firefly luciferase. pLentiEGFPdestablized - EFS-EGFPd2PEST-2A-MCS-Hygro - Lentiviral...Lentiviral vector expressing destabilized EGFP. Lenti-luciferase-P2A-Neo - Lentiviral reporter vector expressing...

...marker. Looking at Table 1, we can recognize a couple of fluorophores from Figure 1. For example, the chart...brightness of individual fluorophores. Table 1 describes the brightness of a few commonly used fluorophores...efficient (i.e., happen most of the time). Figure 1 shows a graph from the BD® Spectrum Viewer, outlining...Resonance Energy Transfer (FRET).   Figure 1: Emission spectra and respective colors of a few commonly...Brighter fluorophores have a stronger signal. Table 1: Degrees of brightness for a few commonly used fluorophores...blue laser excites at a wavelength of 488 nm (Table 1), which is near a local maximum of PE's excitation...Knowing that the red laser excites at 640 nm (Table 1), while comparing the two fluorophore's excitation...

...Tris-tricine 2.5–40 kDa Tris-tricine 30 V, 1 hour or 100 V, 1–2 hours Good separation, quality, and stability...dive in!      Figure 1: The technical decisions to make when designing a western blot.  Psst!...assay. The most common options are 4 °C overnight or 1–2 hours at room temperature (RT). Most people choose...overnight at 4 °C and their secondary antibody for 1–2 hrs at room temperature, which allows them to reduce...weight of the proteins you’re interested in. Table 1 lists the most common types of gels for SDS-PAGE running...gel chemistry and/or specialty gel options. Table 1: Types of gels  Gel Type Protein sizes Running... Tris-glycine 6–400 kDa Tris-glycine 100 V, 1–2 hours Easy and cheap to handcast Short shelf-life...

...Morgens et al. contains three types of guides: 1) targeting guides (blue), which target all protein-coding..., the toxicity of guides with off-target sites was profiled. Guides with exact or 1-bp mismatch off-targets...) or short (17-18 bp) guides. When comparing the toxicity of guides that have 1-bp mismatch off-target... Figure 1: Summary of the advantages of the new approach to genome-wide CRISPR screen design.   Bassik... in both screens, with the growth screen having a 1% false discovery rate and identifying >88% of previously...characteristics that influence gRNA off-target activity: 1) mismatches closer to the PAM are less tolerated than...Knockout Library are available from Addgene! References 1. Morgens, David W., et al. "Genome-scale measurement...

... C, Wright GJ. Analytical Biochem. 2012 May 1;424(1):45-53....are based on the pTT3 plasmid backbone which is described by Durocher et al. Nucleic Acids Research 2002...showing how AVEXIS works with a detailed protocol description: Avidity based Extracellular Interaction Screening...

... your account. We will notify you by email within 1 business day regarding your account verification. ...Plasmid Gene/Insert PI Kits Kit name Type PI Description High Complexity Golden Gate Assembly Standards...Pooled Libraries Pooled Library name Type PI Description HR700_TP53 Exon Mutation Libraries CRISPR Thorsten...Parkinson's Disease; For target validation and drug design Antibodies Addgene distributes ready-to-use recombinant...materials and intellectual property rights, and provides liability protection for material providers. Addgene...

... Caltech and can help guide your decision. Figure 1: Decision tree for choosing your AAV capsid. Your ...tdTomato Control Boyden 83900 pAAV-mDlx-GFP-Fishell-1 Dlx GFP Control Fishell 99130 pAAV-mDlx-NLS-mRuby2...at the bottom of this page. For more in depth descriptions of each serotype, check out Addgene's blog on...Browse Available PHP.eB AAV ID Name Promoter Description Category PI Controls 28306 pAAV-FLEX-tdTomato... Browse Available PHP.S AAV ID Name Promoter Description Category PI 28306 pAAV-FLEX-tdTomato CAG tdTomato...Browse Available PHP.V1 AAV ID Name Promoter Description Category PI 104052 pAAV-CAG-DIO-EYFP CAG EYFP...Browse Available MaCPNS1 AAV ID Name Promoter Description Category PI 37825 pAAV-CAG-GFP CAG GFP Control...

...~50% Table 1: Nanoblades editing efficiencies of primary cells. Comparing Nanoblades to Existing CRISPR...components into 293T cells. Figure 1: The key components of Nanoblades. Image from Mangeot et al., 2019....loaded with 1-4 different gRNAs. Viral envelope proteins To alter the cell tropism of Nanoblades, different... ~50-70% for these four cell types (Table 1). Additionally, hiPSCs and mouse BM cells still behaved like...mouse-model generation. Key components of Nanoblades Nanoblades are a Cas9 RNP delivery vehicle that’s based...) are expressed to pseudotype Nanoblades. The team found that Nanoblades pseudotyped with a mix of VSV-G...using Nanoblades? Plasmids are available from Addgene. Genome editing primary cells with Nanoblades Primary...