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...Production Titer Quality Control Resources Lentivirus Production Titer Quality Control Resources Overview of...subunit (RPP30) as a control for normalization purposes are being used. Quality Control Mycoplasma The 293T...Addgene's viral production methods and quality control for lentivirus and AAV. Viral...virus preps are titered and subjected to quality control by Addgene before being distributed to customers...production protocols, titering methods, and quality control are described below. AAV Production AAV distributed...probes targeting the ITR elements and an internal control of known titer (protocol modified from Lock et ...performed by droplet digital PCR (ddPCR). Quality Control Addgene ensures high quality viral vectors by optimizing...

... inverse fluorescent biosensor for citrate (Citron/Citroff), mitochondrial or cytosolic High-Performance...proteins. Genetically encoded biosensors can be introduced into cells, tissues, or organisms for long-term... with different binding affinities or negative-control constructs used to characterize the sensor's behavior...differ from the original characterization (e.g. in vitro vs. in vivo, cytoplasmic vs. extracellular vs. organelle-localized... GCaMP6f in PV cortical interneurons under the control of the E2 regulatory element Viral manipulation...calcium indicator, based on C-terminal domain of troponin C Griesbeck Lab Twitch Plasmids Oliver Griesbeck...Calcium Red-, yellow-, or cyan-emitting homoFRET/anisotropy Cameleon calcium sensors Single-color, ratiometric...

...murine) ecotropic retroviral transforming sequence C-CBL, CBL2, RNF55 CBLB Cas-Br-M (murine) ecotropic retroviral...TSHA CGB chorionic gonadotropin, beta polypeptide CGB3, hCGB CGB1 chorionic gonadotropin, beta polypeptide... 1 - CGB2 chorionic gonadotropin, beta polypeptide 2 - CGB5 chorionic gonadotropin, beta polypeptide 5...chorionic gonadotropin, beta polypeptide 7 CG-beta-a, FLJ35403, FLJ43118 CGB8 chorionic gonadotropin, beta... CKLFSF8-V2 CNTF ciliary neurotrophic factor HCNTF CNTFR ciliary neurotrophic factor receptor MGC1774 ...14, CST-17, CST-29 CRH corticotropin releasing hormone CRF CRHR1 corticotropin releasing hormone receptor...CSH2 chorionic somatomammotropin hormone 2 CS-2, CSB, hCS-B CSHL1 chorionic somatomammotropin hormone-like...

...GFP and shRNA under the control of a tet-responsive H1 promoter. Didier Trono 11651 pLVUT-tTR-KRAB 3rd...transgene expression. Contains intron that gives high expression. Didier Trono 12254 pWPI 2nd EF-1alpha driven...Plasmids Resources Lentiviruses, a subset of retroviruses, are some of the most common and useful types...for more variants. Patrick Aebischer and Didier Trono 11795 pLL3.7 3rd Expresses shRNA under mouse U6 ...transgene expression and EGFP coexpression. Didier Trono 17619 EF.CMV.RFP 2nd EF-1 alpha promoter for transgene...expression. Miguel Ramalho-Santos 24129 pULTRA 3rd Bicistronic expression of EGFP and the gene of interest. ...Tet-inducible lentivirus for ORF expression, multicistronic design enables tracking of viral transduction...

...Mehl 212125 pBK-Ma pylRS nitroY/haloY-F5 PylRS M. alvus 3-nitrotyrosine (3-nitro-Y), 3-halotyrosine (3-halo-Y...Huber 225684 pAJE-Ma pylRS nitroY/haloY-F5 PylRS M. alvus 3-nitrotyrosine (3-nitroY) and 3-halotyrosine (3...Mehl 85498 pDule-3-nitroTyrosine (5B) 3NY (5B) synthetase M. jannaschii 3-nitroTyrosine Bacterial TAG Ryan...Mehl 85499 pDule2-3-nitroTyrosine (5B) 3NY (5B) synthetase M. jannaschii 3-nitroTyrosine Bacterial TAG Ryan...Chin 174078 pDule-3-nitroTyrosine (A7) 3NY (A7) synthetase M. jannaschii 3-nitroTyrosine Bacterial TAG Ryan... 174079 pDule2-3-nitroTyrosine (A7) 3NY (A7) synthetase M. jannaschii 3-nitroTyrosine Bacterial TAG Ryan...generally used. Once these plasmids have been introduced in the cell, the ncAA provided in the media can...

...regions controlling transcription Debrya Groskreutz 18760 MSCV IRES Luciferase Firefly Retroviral expression...contain introns Anthony Leung Return to Top Expression Constructs Luciferase driven by a strong constitutive...luciferase gene fusions. Renilla luciferase under the control of a CMV promoter is present for normalization ...RapidResponse™ Renilla Vector for investigating regions controlling transcription Pete Stecha 212933 pGL4.82(hRluc...Puro) Renilla Vector for investigating regions controlling transcription Pete Stecha 106292 pLS-SV40-mP-...fusions in plants. Renilla luciferase under the control of a CMV promoter is present for normalization....reporter reporter vector allowing the study of biscistronic promoters. Daniel Christophe 119760 pSGDlucV3.0...

...Boyden Voltage Reporter: Voltron 119036 pAAV-hsyn-flex-Voltron-ST Syn Voltron-ST none Cre dependent 1 ...filtering options. *AAVrg = retrograde serotype, produced with the AAV retro helper plasmid from the Karpova...proteins, including calcium sensors, under the control of various promoters. Sensor Green Calcium Sensors...Oxytocin (OT) Sensors GRAB_OT Voltage Sensors Archon Voltron JEDI-2P ASAP ATP Sensors iATPSnFR2 5-HT Sensors... dependent) Serotype AAV1 AAV2 AAV5 AAV8 AAV9 Retrograde AAV PHPeB Clear Filters ID Name Promoter Sensor...pAAV-hSynapsin1-axon-GCaMP6s-P2A-mRuby3 Syn axon-GCaMP6s bicistronic mRuby3 Constitutive 5, 9 Lin Tian 112010 pAAV-hSynapsin1... 140555 pAAV-hsyn-GRAB_DA-mut Syn GRAB_DA-mut (control) none Constitutive 9 Yulong Li 140556 pAAV-hsyn-GRAB_rDA1m...

...CRISPR Libraries Cas9 Pooled Barcoding Libraries Control Addgene's lentiviruses are made from plasmids that...76,441 unique sgRNAs along with 1000 non-targeting controls. This backbone does not contain SpCas9. 76,441...,114 human genes along with 1000 non-targeting controls Puromycin Doench , Root 73179 Human gRNA pooled...76,441 unique sgRNAs along with 1000 non-targeting controls. This backbone contains SpCas9. SpCas9 and 76,...,114 human genes along with 1000 non-targeting controls Puromycin Doench , Root Human activation pooled...57,050 unique sgRNAs along with 500 non-targeting controls. This backbone does not contain dCas9. 57,050 ...18,901 human genes along with 500 non-targeting controls Puromycin Doench , Root 92386 (Discontinued) Human...

... capacity. Gamma-retroviral Vector Guide | Gamma-retroviral Plasmids Gamma-retroviral vectors, often referred...Guides and Plasmids Selecting a Viral Vector Type Introduction to Viral Vectors Resources Viral vectors are... referred to as just retroviral vectors, have a relatively high transduction efficiency and only infect...genome, enabling long-term transgene expression. Retroviral vectors are popular for making stable cell lines...transient transgene expression.They can induce a strong immunogenic response, useful for clinical applications...80–130 nm Dividing/Non-dividing Yes Low Gamma-retroviral Stable RNA < 8 kb 80–130 nm Dividing Yes Moderate...frequency of target cell genome integration. Introduction to Viral Vectors Figure 1: Wild-type virus propagation...

...facilitation, DNA amplification, and high-quality quality control measures such as titering, purity assessment, sterility...is used in a wide range of experiments, both in vitro and in vivo. It has low immunogenicity, is easy ...serotype? Learn more about AAV serotypes and tissue tropism on Addgene's blog . All-in-One Pricing The price...centrifugation purification Addgene’s comprehensive quality control process: Titering Purity assessment Sterility testing...produce the prep, and perform comprehensive quality control processes to make sure it is ready for your experiments...your AAV vector experience, from methods to troubleshooting insights, you make research more reproducible...gap between published methods and real-world troubleshooting. Eligible Plasmids The following plasmids are...

...81070 rAAV2-retro helper AAV retrograde AAV packaging plasmid, expressing Rep2 and retrograde capsid Alla... transgene or genetic elements of interest to introduce in cells. A packaging plasmid (or Rep/Cap plasmid...non-standard), expressing Rep2 and PHP.eB Cap controlled by tTA-TRE amplification system Viviana Gradinaru...(non-standard), expressing Rep2 and PHP.S Cap controlled by tTA-TRE amplification system Viviana Gradinaru...non-standard), expressing Rep2 and PHP.V1 Cap controlled by tTA-TRE amplification system Viviana Gradinaru...plasmid (non-standard), expressing Rep2 and B10 Cap controlled by tTA-TRE amplification system Viviana Gradinaru...plasmid (non-standard), expressing Rep2 and B22 Cap controlled by tTA-TRE amplification system Viviana Gradinaru...

... undergo stringent quality control by Addgene or a trusted quality control partner before distribution...are initially titered by spectrophotometry using a NanoDrop spectrophotometer. If needed, the sample is...Addgene's antibody production methods and quality control processes for purified recombinant antibodies. ...production protocols, titering methods, and quality control are described below. Information for a specific...samples with >90% purity are distributed. Quality Control Each lot of antibody undergoes testing, either ...fluorescence pattern is compared to an untransfected control. Immunohistochemistry (IHC) Tissue sections are...Protein expression is compared to an untransfected control and the protein size is confirmed. Residual Plasmid...

...visualization, study of protein interactions, controlled gene expression, reporter plasmids and genome...Purification Protein Visualization Protein Interactions Controlled Expression Reporter Plasmids Genome Engineering...arrays of zinc finger nucleases. Addgene Blog Introduction to FRET Bimolecular Fluorescence Complementation...Studying Protein–Protein Interactions Return to top Controlled Expression It isn’t always advantageous to constitutively...stationary phase. In these cases, you may want to control protein levels or to turn on expression only at...contains plasmids whose expression levels can be controlled by a variety of small molecules, light, and temperature...Escherichia coli Yingfeng An Additional Addgene Controlled Expression Resources Check out our Tetracycline...

...systems, how to use them to modify the genome or control gene expression, and find plasmids for your experiments...engineering tools used to precisely manipulate DNA and control the expression of specific genes in cells or organisms...recombinases can be used to modify the genome or control gene expression or skip ahead to browse highlighted...Summary of Cre, Dre, and Flp Recognition Sites. Controlling Recombinase Expression By expressing the enzyme...promoters or inducible systems, you can precisely control and optimize expression of your gene of interest...fused to a ligand-binding domain such as the human estrogen receptor (ERT2) so that the enzyme is only activated... and C-terminal fragments and placed under the control of different promoters. Expression of both fragments...

...telangiectasia Didier Trono 14581 pSUPER.ATMi ATM H1 Ataxia telangiectasia Didier Trono 14666 pQE9-His-p97...T7 Parkinson's Dmytro Yushchenko 107293 s2. pT7-7-SNCA-5Asp SNCA T7 Parkinson's Dmytro Yushchenko 107294... T7 Parkinson's Dmytro Yushchenko 107295 r. pT7-7-SNCA-5Asn SNCA T7 Parkinson's Dmytro Yushchenko 107296...T7 Parkinson's Dmytro Yushchenko 107297 r-22. pT7-7-SNCA-9Asn SNCA T7 Parkinson's Dmytro Yushchenko 107425...neuroaxonal dystrophy Michael Ward 178167 RNF216_Halo_C_allele RNF216 Halo Hypogonadotropic hypogonadism...Ataxia Pietro De Camilli 194005 VPS13D1-1576-(Y1253S) VPS13D GFP CMV Spinocerebellar Ataxia Pietro De Camilli...Parkinsonism Pietro De Camilli 232865 mCherry-VAB(VPS13C) VPS13C mCherry CMV Parkinsonism Pietro De Camilli...

...Image from Fenno et al., 2014. INTRSECT INTRSECT (intronic recombinase sites enabling combinatorial targeting...and Flp) recognition sequences within synthetic introns. INTRSECT was first shown as a proof-of-concept...transgenic recombinase-expressing animal lines and retro-grade expressing viruses delivering additional recombinases.... Figure 2: INTRSECT works by inserting short, intronic sequences into the open reading frame (ORF) of...sequences (e.g. Lox sites, FRT sites) inside of the introns (A,D). The addition of these recombinase recognition...recognition sequences enables directional control of the portion of the ORF that is sandwiched between the... combinations of recombinases are present, the introns are excised during mRNA processing, producing a...

...PGKpuro2ABFP - For retroviral delivery of one sgRNA pMKO.1 GFP - Retroviral shRNA expression Retroviral backbones...test the function of your gene of interest in a controlled environment under various conditions. When choosing...tag for mammalian expression (Gateway) pYIC - Bicistronic fluorescent reporter gene with cap-dependent ...expression, see our dedicated Lentiviral Plasmids page Retroviral Easy and safe to use, integrate into host genome...MSCV-IRES-GFP or pMSCV-IRES-mCherry FP - Mammalian retroviral gene expression with GFP or mCherry selectable... luciferase vector for investigating regions controlling transcription See our Luciferase collection of... production AAV-GfaABC1D-MCS--4x6T-WPRE - For astrocyte-selective expression in AAV Dual promoter Separate...

...be measured using fluorescence microscopy or spectroscopy by exciting the donor and detecting the emission...available, we have listed cloning backbones and control plasmids. Donor Acceptor λ Dex QY D λ Aem ε A QY...refer to published reports and include appropriate controls to help interpret your results. See Algar et al...feature plasmids with positive and negative FRET controls or calibration standards consisting of donor and...quantitative two-hybrid FRET Positive and negative controls and cloning backbones for mTurquoise2-mVenus(L68V...towards a quantum yield of 93% Positive and negative controls and cloning backbones for various FRET pairs with...acceptors for mTurquoise2 Positive and negative controls and cloning backbones for red, yellow, and turquoise...

...Ready-to-use AAV available from Addgene's viral service. Control EGFP vectors in various serotypes for serotype ... AAV2, AAV5, AAV6, AAV8, AAV9, AAV11, and AAV retrograde....fluorescent reporters and can be used to compare the tropism of different serotypes. In addition to the standard...list of all 100 µL sized sample, please see our Control AAV Inventory . The catalog ID for the 20 µL sized... catalog item, including the titer and quality control, visit the material page (linked for each catalog...AAV11 37825-AAV11.T 20 µL $ 150 Add to Cart AAV Retrograde 37825-AAVrg.T 20 µL $ 150 Add to Cart pAAV-hSyn-EGFP...

...provide a basic understanding of CRISPR biology, introduce the various applications of CRISPR, and help you...research. CRISPR Basics In this section, we will introduce the basic mechanisms of CRISPR and some of the... nuclease fidelity SpCas9-NG — NG; increased in vitro activity SpG — NGN; increased nuclease activity ...-occurring Acr (Anti-CRISPR) proteins can help control CRISPR activity. Acr proteins have evolved in phages...LmoCas9 and SpCas9. Acr proteins can be used to control both when and where genome editing happens. CASANOVA...Dominik Niopek’s lab , is a way to optogenetically control genome editing by fusing the anti-CRISPR protein...photosensitive LOV2 domain . Several labs have also controlled Acr activity with endogenous cell-type specific...