...journal. An increasing number of funding bodies insist that the research data resulting from their funding...many journals use to verify whether a manuscript contains all the necessary information to be of maximal...

...interested in following? Someone you admire for their insight and way of expressing it in meetings? The more ...help you learn the local “ropes” and become an insider faster A mentor that works elsewhere is able to...

...biomedical researcher at Lunenfeld-Tanenbaum Research Institute, posting about Addgene’s 20th anniversary on Twitter...alike. We transformed this process by aligning institutions with standardized agreements and electronic ...

... for packaging, however, the isoforms of these proteins, as well as the viral LTRs differ. Therefore, ...Retroviral Barcoding Library (Kit # 1000000097) The Winslow lab barcoding library consists of 96 retroviral...

... a STAR answer once the interview has started. Instead, I preferred to prepare stories to answer interview...possible. Otherwise, read the video conferencing instructions you received from HR carefully. And even if ...

.... The advantages provided by chemogenetics are instead that it is noninvasive and has longer-lasting effects.... PSAMs themselves are modified ligand binding domains. When coupled to the ion pore domain (IPD) of a...

...example: in a cell there are proteins which interact with other proteins and with DNA, and these complexes...

...in a variety of outcomes including excision or insertion, inversion, translocation or cassette exchange...addition, Casɸ has a minimal PAM requirement. For instance, for Casɸ-2 the PAM sequence is 5’-TBN-3’ (where...

...last digit: An answer should come soon, but not instantly. Addgene tries to get back to customers within... Eric J. Perkins...

... now specifically drives expression of mCherry instead of BFP. Beyond switching fluorophores, why is FLEx...optogenetic reporter would ensure that the reporter remains silent until a cell or tissue-specific Cre is provided...

...agarose/DMEM solution against a part of the body very sensitive to heat- the inside of the wrist or the ...

...deposition of the fertilized egg by its mother. Inside the pupal case, the larva will metamorphose into...isolate females from males while they are still virgins. Females begin mating ~6 hours after hatching into...

...the looping of chromatin, the complex of DNA and proteins that forms chromosomes. These long range DNA interactions...Next Experiment Learn More About Designing gRNAs Against Your Favorite CREs Here Additional Resources on...

...questions. Practice your presentation for both instances. If your poster material is nearing publication...Connect with people on LinkedIn or Twitter, for instance. If you’ve had a particularly meaningful conversation...

...plasmid of interest, for detecting the presence of an insert or piece of DNA in a sample such as in colony PCR...into approximately 20,000 droplets. Each droplet contains the material required for PCR amplification. Following...

...widely used method due to the accessibility of instruments in standard labs. When determining titers by ...., Pawlita, M., Ferrari, F., Samulski, R., & Kleinschmidt, J. (1999). Titration of AAV-2 particles via...

...light to distinguish one fluorophore from another. Instead of using a single (primary) detector to identify...narrow band, to discern one marker from another. Instead of compensation, which relies on user adjustment...

...transcription and chimeric efforts make a precise edit or insertion via a chemical modifier. We used these categories... plasmids seem to never truly go out of style. Instead, an almost even mix of plasmids (by age) are requested...

...including casual request via email, in person, or instant messaging but we as scientists often prefer data...failing in many academic journal clubs where institutional inertia drives the meeting rather than interest...

...information. Personally, I would much rather read the instructions than watch a how-to video. My dad, on the other...love it and others will avoid it like the plague. Instead of looking for The One Method to Rule Them All,...

...time to produce beta-lactamase prior to division, instead of immediately experiencing the effects of the ... capable of correctly folding disulfide bonded proteins in its cytoplasm. Microbial Cell Factories, 11...

...Eleven years later,  Addgene is a company of fifty instead of three, and we are sending hundreds of plasmid...yard this year, though she generously offered. Instead we picniced outside of our new office space on ...

... vector (deposited by David Root of the Broad Institute) has consistently been one of the repository’s...satisfaction of checking off each step as you painstakingly complete your experiment – one small victory...

...run and the long run and so will your work. Nobody wins, and you need to make a change. #2 Thou shalt sometimes...tasks – try to resist the impulse to control by insisting things always be done your way. Be patient with...

...Livny J, Regev A, Koonin EV, Hung DT, Sabeti PC, Collins JJ, Zhang F (2017) Nucleic acid detection with ...Gootenberg JS, Abudayyeh OO, Kellner MJ, Joung J, Collins JJ, Zhang F (2018) Multiplexed and portable nucleic...

...open new ways for a novel therapeutic approach against drug addiction. Anxiety tests Rodents have a natural...are ahead for behavioral neuroscience to reveal insights into the complex relationship between genes, neurons...

...is not amenable to FACS.  Thus, the researchers inserted human CD8, a gene that encodes a surface protein...yourself whether gene overexpression is necessary (instead of gene knockout or knockdown, for which there ...

...manipulated by injected RNA and can express fluorescent proteins. This lab has created an automated tank for feeding... fusion is such that the two plants share RNA, proteins, and DNA. Euprymna scolopes (Joshua Rosenthal...

...for scientists making this transition. In this installment of the Addgene career guide, we’ll think about... Eric J. Perkins...

...you can either order a DNA pooled library, which contains a collection of plasmids in a single tube, or ...lentiviral CRISPR libraries. Protocols from the Broad Institute- A variety of protocols and guides regarding pooled...

...circuit is sufficient to induce reinforcement (instills a sense of “reward”), then the animal will repeatedly...derived from direct comparative analysis of microbial opsins." Nature methods 9.2 (2012): 159-172.5. Goshen ...

...the mission so much that they will often move mountains to give advice and guidance to those that ask....great sources of volunteer opportunities. NSPN maintains a great resource guide including conferences, ...

...GFP CMV Parkinson's L. Miguel Martins 14122 pcDNA3-HtrA2-EGFP S306A HTRA2 GFP CMV Parkinson's L. Miguel...Miguel Martins 14124 pcDNA3-delta133HtrA-EGFP HTRA2 GFP CMV Parkinson's L. Miguel Martins 14126 pET20b-delta133HtrA2...Miguel Martins 15939 pcDNA3-HtrA2-FLAG S142D HTRA2 Flag CMV Parkinson's L. Miguel Martins 15940 pcDNA3-HtrA2...Flag CMV Parkinson's L. Miguel Martins 16153 pET20b-delta133HtrA2 S142A HTRA2 His T7 Parkinson's L. Miguel...Miguel Martins 16154 pET20b-delta133HtrA2 S142D HTRA2 His T7 Parkinson's L. Miguel Martins 16155 pET20b-...His T7 Parkinson's L. Miguel Martins 16156 pET20b-delta133HtrA2 S400D HTRA2 His T7 Parkinson's L. Miguel...Miguel Martins 16157 pET20b-delta133HtrA2 A141S HTRA2 His T7 Parkinson's L. Miguel Martins 16158 pET20b-...

...inactive “dead” Cas9 (dCas9) contains mutations in both nuclease domains, D10A and H840A, resulting in...naturally-occurring Acr (Anti-CRISPR) proteins can help control CRISPR activity. Acr proteins have evolved in phages ...bidirectional insertions — meaning they may not be suitable for applications where orientation of the insert is ...exhibit almost unidirectional insertions but are more prone to off-target insertions. And in the case of both... edits. These proteins, like Cre recombinase or phage derived serine integrases, insert donor DNA at “...used to install the LSR Bxb1’s landing site into the genome at a desired locus, then Bxb1 installs the donor...and antibody-activator effector proteins; long-term imaging of proteins in living cells (Figure 9B) dCas9...

...four protein chains: two large heavy chains and two small light chains. These four domains come together...their monovalent nature. Darpins - Darpins are genetically engineered small proteins derived from ankyrin ...ankyrin repeat proteins. These are antibody mimics instead of modified antibodies and are not closely related...synthetic binding proteins which use the Fibronectin type III domain as their scaffolding instead of the antibody...Denature proteins, using heat and/or chemicals, and run on an SDS-PAGE gel to separate proteins by size...your antibody’s specificity. A blank (contains water or buffer instead of a biological sample), which captures... lab to specifically and tightly bind to other proteins. Antibodies, produced as part of the immune system...

...Firefly Insertion of 5' promoter/enhancer regions Bert Vogelstein 12178 pIS0 Firefly Insertion of target... Renilla Insertion of target sequence in the 3' UTR David Bartel 12177 pIS2 Renilla Insertion of target...improved normalization. John Atkins 167576 mpCHECK2 Firefly/Renilla Insertion of 3' UTR sequences downstream...construct that contains your regulatory element of interest. ID Plasmid Description Gene/Insert PI Return ...Plasmid Collection You may also like... Fluorescent Proteins Blog: Luciferase 101 Blog: Technologies Enabled...organisms, ranging from bacteria to aquatic animals to insects. Luciferase catalyzes the oxidation of a substrate...elements on gene expression. Empty backbones for inserting regulatory elements before or after the luciferase...

... E MGC4638 INS insulin ILPR, IRDN INS-IGF2 INS-IGF2 readthrough transcript - INSL3 insulin-like 3 (Leydig...MGC119819, RLF, RLNL INSL4 insulin-like 4 (placenta) EPIL, PLACENTIN INSL5 insulin-like 5 MGC126695, MGC126697...MGC126697, PRO182, UNQ156 INSL6 insulin-like 6 RIF1 INSR insulin receptor CD220, HHF5 JAG1 jagged 1 (Alagille...communication between cells of the immune system. Interleukins Interleukins are cytokines that are secreted by immune... omega 1 - IGF1 insulin-like growth factor 1 (somatomedin C) IGF1A, IGFI IGF1R insulin-like growth factor...MGC18216 IGF2 insulin-like growth factor 2 (somatomedin A) C11orf43, FLJ22066, FLJ44734, INSIGF, pp9974 IGF2R...bA12D24.1.1, bA12D24.1.2 RLN3 relaxin 3 H3, RXN3, ZINS4, insl7 RNASE2 ribonuclease, RNase A family, 2 (liver...

...Plasmids 101: E. coli strains for protein expression Plasmids 101: Common Lab E. coli Strains Why Bacteria? E...Photoswitchable fluorescent proteins Various PIs Fluorescent protein fusion: contains empty vectors with N and...Expression: contains mEos2 with an N-terminal His tag. Photoactivable fluorescent proteins like this one... 3OC6HSL (Quorum sensing molecule) Cynthia Collins Contains luciferase inducible by the quorum sensing...factor bind to a particular sequence? Do these two proteins interact? What happens when I overexpress this...From recombineering to CRISPR , this collection contains a variety of tools to help you harness bacterial...binding sites Philippe Cluzel RNA localization: Contains a 2x array of the binding sequence for RNA-binding...

...utilized CRISPR as a powerful defensive strategy against viral invaders, and this system is proving to be...genome in 1987, but their function as a safeguard against bacteriophages was not elucidated until 2007. Scientists...re-exposure. More specifically, specialized Cas proteins snip foreign DNA into small fragments approximately...stretches of DNA known as CRISPR arrays. Separate Cas proteins then express and process the CRISPR loci to generate...system. Within the bacterial genome, a CRISPR array contains many unique protospacer sequences that have homology... potential target sequence is only valid if it contains a special Protospacer Adjacent Motif (PAM) directly... varying PAM sequences and number/types of Cas proteins. Makarova et al. ’s classification defines 5 types...

...loops at tetraloop and stemloop - Contains BbsI sites for insertion of guide sequences Addgene plasmids...2 and EF1a-zeo resistance marker. Contains BsmBI sites for insertion of guide sequences Zhang Lab SAM ... microbial nuclease system involved in defense against invading phages and plasmids. CRISPR loci in microbial...nickase) + CRISPR RNA array + tracrRNA: This plasmid contains three expression cassettes. To target a given ...NGG PAM sequence. 3. SpCas9 alone: This plasmid contains a single cassette expressing human codon-optimized...2. SaCas9 only: This plasmid ( PX600, #61592 ) contains a single cassette expressing human codon-optimized...SpCas9 Cas9: The PX551 plasmid (AAV-SpCas9, #60957) contains a single cassette expressing human codon-optimized...

...reverse transcriptase (RT) is capable of installing targeted insertions, deletions, and all possible base-to-base...joining (NHEJ), a mechanism which frequently causes insertions or deletions (indels) in the DNA. Indels often...repair a DSB using homology-directed repair (HDR) instead of NHEJ. In most experimental systems, HDR occurs...much lower efficiency than NHEJ. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Browse...mutants for directed evolution. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Nick ...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type...homology-directed repair (HDR). ID Plasmid Gene/Insert Promoter Selectable Marker Publication Prime Edit...

...mCherry tagged fusion proteins in worms ins:MCS; cryaa:RFP - A zebrafish insulin promoter vector with ...designed for a user to insert a gene of interest. They may contain tags or fusion proteins, a multiple cloning...and pFastBac Dual LIC vectors for insect expression Fluorescent proteins (GFP, mCherry, etc) Localization...molecular biology to isolate, multiply, or express the insert they carry in the target cell. These vectors allow...function section for if you want to stably express an insert using viral vectors, are studying a different genetic... - Inducible expression, fusion proteins, and more pGBKT7-GW - Gateway compatible Y2H vector...vector pFA6a vectors - Epitope tagging in S. pombe Insect/Baculovirus UAS, MT, Polyhedrin Targeted Gene ...

...microbial opsins, such as the Halorhodopsins, Archaerhodopsins and Leptosphaeria rhodopsins, are pumps...encode proteins that are controlled by light. Many of these plasmids encode for microbial opsins, which... opened and closed in response to light. These proteins can be expressed in specific neurons, often utilizing...in a precise manner. One of the first microbial opsins to be adopted as an optogenetic tool is the widely...anions into or out of the cell. When using these opsins as a tool to control neuronal activity, scientists...expression and localization. Many other types of proteins can also be controlled by light, generally by ...phytochromes, or light-oxygen-voltage-sensing (LOV) proteins. These fusions can then be used as an optical ...

...commonly used adenovirus terms Adenoviral Strains Bacterial strains for generating adenovirus. ID Strain Description...beta-gal Shuttle Test plasmid that contains β-gal and GFP; contains ∼10Kb genomic DNA stuffer Vogelstein...modular inserts for seamless assembly of multiple mutations, alterations, or transgene insertions into single...Collection of adenovirus shuttle vectors and strains Viral...Vogelstein 16399 AdEasier-1 BJ5183 derivative that contains AdEasy-1™ plasmid Vogelstein Adenoviral Plasmids...database for adenoviral transfer plasmids that have inserts. ID Plasmid Type Description PI 16400 pAdEasy-1...recombining shuttle plasmids; for use with large inserts Vogelstein 179201 pAd5-B6 ∆E3-GFP Adenoviral A ...

... monoclonal antibodies against zebrafish neural receptors and secreted proteins suitable for wholemount...of membrane proteins. Elife. 2018 May 24;7. and Generation of synthetic nanobodies against delicate proteins...Antibodies Antibody Plasmids Antibodies are large proteins made by the immune system that bind to an antigen...pull out a protein of choice from a mix of other proteins, visualize a protein under a microscope, or detect...in bacteria, mammalian cells, yeast, plants, or insect cells. Benefits include consistency between lots... Antibodies: Selecting and Evolving Functional Proteins. METHODS: A Companion to Methods in Enzymology...to rapidly engineer the constant immunoglobulin domains to obtain recombinant hybridomas, which secrete...

...reverse transcriptase (RT) is capable of installing targeted insertions, deletions, and all possible base-to-base...joining (NHEJ), a mechanism which frequently causes insertions or deletions (indels) in the DNA. Indels often...repair a DSB using homology-directed repair (HDR) instead of NHEJ. In most experimental systems, HDR occurs...much lower efficiency than NHEJ. ID Plasmid Gene/Insert Promoter PI Publication Browse CRISPR Plasmids ...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type...homology-directed repair (HDR). ID Plasmid Gene/Insert Promoter PI Publication Prime Edit Cas9 H840A nickase...desired edits on an RT template. ID Plasmid Gene/Insert Promoter PI Publication Activate Catalytically ...

...or MCP tagged plasmids for visualization of proteins inside cells....-, ACP-, and MCP-tags for visualization of proteins inside living or fixed cells. The plasmids associated... labeling cell-surface proteins, and are useful for labeling secreted proteins with disulfide bridges ...features: Allows for visualization of multiple proteins simultaneously with dual labeling when used with...live or fixed cells, and for internal or surface proteins Single constructs are compatible with multiple... and MCP-tags are used exclusively for surface proteins and use enzymatic binding to attach substrate ...suitable for both cell-permeant and non-cell-permeant proteins. CLIP-tag CLIP-tag is identical in function to...

...correct insert (gRNA) in pX330 / pX335 with BbsI/AgeI double digestion. As a successful insertion will destroy... have been published with thorough step-wise instructions on designing and using Cas9-based genome engineering...vector (like PX330). For large changes (>100bp insertions or deletions), we typically use plasmid donor...homology arms on each side flanking your desired insertion or mutation. Each arms is around 800bp. For transfection...CRISPR/Cas9 system to create specific mutations or insertions by Homologous Recombination (HR), what is the...purified long oligos. For large changes (>100bp insertions or deletions), the Zhang lab typically uses a...homology arms on each side flanking your desired insertion or mutation. Each arm is around 800bp. For transfection...

...Mullineaux Insulin Sensor for Imaging Insulin Secretion A Ratiometric Sensor for Imaging Insulin Secretion... Collections Fluorescent Proteins Fluorescent Biosensors Fluorescent Proteins: Biosensors Metal Ions Small...Service: AAV Biosensors Fluorescent biosensors are proteins designed so that their fluorescent properties ...transfer (FRET) efficiency of a pair of fluorescent proteins. Genetically encoded biosensors can be introduced...Constitutive or Cre-dependent) Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature. 2013 Jul...nested reporters of green- and orange-emitting proteins Ratiometric Matryoshka biosensors from a nested...cassette of green- and orange-emitting fluorescent proteins. Nat Commun. 2017 Sep 5;8(1):431. Wolf Frommer...

... EF1a-tTA-2 Contains Gateway L1L3 sites and EF1α promoter driving tTA; contains insulator sequence; plasmid...Blasticidin selection. Please NOTE, this vector contains an insert (MAT2A) which would need to be replaced by...Backbones Find a construct that will allow you to insert and induce your gene of interest. ID Plasmid Description...format: PGK-rtTA-2A-puro; see article for tagged insert options rtTA On Root 11651 pLVUT-tTR-KRAB Lentiviral...bidirectional tet-responsive promoter (Pbi); Pbi contains a TRE between two minimal CMV and is silent in...third-generation rtTA On Elledge 19407 pTREtight2 Promoter contains a modified TRE that is silent in absence of binding...