...and easiest way is to co-transfect several plasmid together, with each plasmid having a targeting spacer... may also like... Zhang Lab CRISPR plasmids General CRISPR Plasmids Dr. Le Cong, the first author of one... each locus into PX330, then co-transfect or delivery the two plasmids together into your cells. If you... For transfection in cells like HEK, we use around 400ng of a typical size (~ 5kb) donor plasmid with ...positively transfected. Alternatively, you can use a selection marker to select transfected cells (for...example, plasmid with a puromycin resistance cassette, such as PX459 ). After you co-transfect the CRISPR... Genome Engineering CRISPR Zhang Lab CRISPR Plasmids Zhang Lab CRISPR FAQs Zhang Lab's CRISPR Frequently...

... Cas9 plasmid Bullock and Port 51026 U6-BbsI-crRNA dU6 BbsI Transfection none, need Cas9 plasmid O'Connor-Giles...CRISPR Plasmids: Drosophila Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime...gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter...with a wide variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance... none, need Cas9 plasmid Bullock and Port 49330 pAc-sgRNA-Cas9 dU6 BspQI Transfection Puromycin yes, cut...Liu 45946 pU6-BbsI-chiRNA dU6 BbsI Transfection none, need Cas9 plasmid O'Connor-Giles, Harrison, Wildonger...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...

...usage of the pSpCas9(BB) plasmid (pX330) (Addgene plasmid ID 42230). This plasmid allows for the simultaneous...another transfection method such as cationic liposome-based transfection. Optimize transfection conditions...utilized, such as pX458 (Addgene plasmid ID 48138) or pX459 (Addgene plasmid ID 48139), which include GFP ...for MEL cells. However, transient transfection of CRISPR/Cas9 plasmids may be successfully adapted to ... with a PCR cloning kit) into a plasmid vector. Transform the plasmid into DH5α E. coli cells and plate...Mammalian Cell Lines Other Addgene Resources CRISPR Plasmids CRISPR Guide Addgene Blog Posts Addgene is proud...multiple sgRNAs may be expressed from a single plasmid. Add “CACC” before the 20-mer guide sequence and...

...List of plasmid kits and accessory plasmids available from Addgene for generating your own specific TALE...Yamamoto Additional Plasmids for Use with Addgene TALEN Kits For Use With: ID Plasmid Depositing PI Description...destination plasmids were optimized for TALEN expression in mammalian systems including hard-to-transfect cell...TALE Toolbox 49622 - 49647 24 plasmids Thoru Pederson TALEColor plasmids: destination vectors for use ...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail... TALE plasmids. Genome... Genome Engineering TALENs TALEN Plasmids and Kits You may also like... Addgene's Genome...

...Grade DNA (cgDNA) for over 200 plasmids in our collection. The plasmids available in this format include...variety of popular plasmids and backbones with high cloning potential. By making these plasmids available as...individual plasmid page to find current pricing information Standard Quality Control Plasmid verified by...next generation sequencing Plasmids Available as Cloning Grade DNA ID Plasmid Description Industry PI FAQ...grade DNA preps of this plasmid available, but helps to inform us of which plasmids would be most helpful... our most popular plasmids! DNA... DNA Service: Cloning Grade DNA Service Details Plasmids Available FAQ Addgene...

...For easy-to-transfect cell types (e.g. HEK293 cells), transfection with standard transfection reagents may...Additionally, many plasmids containing validated gRNAs are now available through Addgene. These plasmids contain ...particular model organism. There may not be a perfect plasmid for your specific application, and in such cases...different activators exist, including the multi-plasmid SAM system. Select Expression System To use CRISPR...to generate stable cell lines (for details, see Plasmids 101: Mammalian Vectors ) Transient or stable expression...or gRNA in a mammalian cell line that can be transfected at high efficiency Lentiviral transduction Cas9...and enrich positive cells Packaging and envelope plasmids provide the necessary components to make lentiviral...

...Cas9 plasmid Joung 46170 PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9 plasmid Calarco...gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter...with a wide variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance...Calarco 47943 pMB70 cU6 BsaI Transfection none, need Cas9 plasmid Boxem 47941 pMB60 T7 BsaI In vitro transcription...elegans none, need Cas9 plasmid Ristow Do you have suggestions for other plasmids that should be added to this...CRISPR plasmids that have been designed for use in C. elegans. CRISPR... CRISPR C. elegans CRISPR Plasmids: C. elegans...

...Collections Luciferase Plasmids Luciferase Plasmid Guide You may also like... Fluorescent Proteins Blog: Luciferase...luciferase. Plasmids are Gateway cloning compatible. Learn more about DULIP in our 27 Hot Plasmids from 2016...luciferase reporter. These plasmids improve over the original STARR-seq plasmids by using the bacterial ORI...otherwise noted, these plasmids are meant for expression in mammalian systems. ID Plasmid Luciferase Type(s...luciferase expression plasmids; these are noted with the viral service icon . ID Plasmid Luciferase Type Promoter...AAV9-CRISPR knockout plasmid. Renilla luciferase is used to monitor viral transfection Feng Zhang 21471 pLenti...AAV-CRISPR knockout plasmid. Firefly luciferase is used to moniter viral transfection Sidi Chen 109396 TBG-Vector...

... low levels of co-purified plasmid from transfection. This residual plasmid is extracted and amplified...vectors, human embryonic kidney cells are transfected with the plasmid(s) encoding the heavy and light chains...compared to an untransfected control and the protein size is confirmed. Residual Plasmid Sanger Sequencing...the Sanger method, and aligned with the parental plasmid sequence to confirm the antibody’s identity. For... The fluorescence pattern is compared to an untransfected control. Immunohistochemistry (IHC) Tissue sections...

... Plasmid Collections EXPRESs Plasmids Zebrafish Monoclonal Antibodies Zebrafish...also like... Gavin Wright Lab plasmids This is a collection of plasmids that encode monoclonal antibodies...BMC Biol. 2010 Jun 4;8:76. PubMed 20525357 Plasmid ID Plasmid Name Zebrafish Gene 28214 SI1-Unc5b Unc5b...This is a collection of plasmids that encode monoclonal antibodies to zebrafish cell surface receptors...cell surface receptors and secreted proteins. The plasmids encode both the light and heavy chains of mouse... proteins and the antibodies are produced by transfecting mammalian cell lines and collecting the secreted...

...) via transfection, or shRNA (short hairpin RNA) via transfection or viral delivery of a plasmid. Below...Plasmids for mammalian RNA interference (RNAi) experiments at Addgene; browse by constitutive lentiviral... Plasmid Collections Mammalian RNAi Mammalian shRNA Tools... libraries Lentiviral packaging and expression plasmids Background RNA interference (RNAi) is an RNA-mediated...selection of empty vectors for cloning in shRNAs. Plasmids for constitutive expression as well as those that...inducible (Tet) expression are available. To find plasmids containing RNAi components, like Argonaute, please...Lentiviral vectors for constitutive shRNA expression ID Plasmid Description PI 10878 pLKO.1 ‐ TRC cloning vector...

...tracrRNA: This plasmid contains three expression cassettes. To target a given site, the plasmid can be digested...using SAM: Addgene plasmids #61422 - #61424 were designed for transient transfection : 61422 : Expresses...adenocarcinoma pathology. These plasmids as well as a backbone plasmid for cloning new targets are described...sgRNA backbone for cloning new targeted plasmids. The plasmid can be digested using SapI, which will reveal...2015 Apr 1. PubMed . Return to AAV In vivo plasmids AAV Plasmids with Cas9 Mouse CRISPR-Cas9 knockin mice...

...Table 2. pmax dCas9 Activator Expression Plasmids ID Plasmid 48223 pAC91-pmax-dCas9VP64 : dCas9VP64 on...CRISPR Cas9 gRNA plasmids from Dr. Rudolf Jaenisch's lab. CRISPR... CRISPR Jaenisch Lab CRISPR Plasmids Jaenisch Lab...Lab CRISPR Plasmids Available from Addgene You may also like... CRISPR Guide CRISPR Protocols gRNA Design...dCas9 activators and sgRNAs can be separately transfected (pmax dCas9 activators [48223~48227] + sgRNA ...different systems by LR clonase reaction. These plasmids are described in: Multiplexed activation of endogenous...Link opens in a new window) Article . Individual plasmids can be ordered via the links below: Table 1. dCas9...

... Viral Plasmids Adenovirus Plasmids Adenovirus Plasmids Additional Resources: Adenovirus...yeast. Two plasmids, termed a shuttle plasmid and an adenoviral (also called backbone) plasmid, are recombined...contains AdEasy-1™ plasmid Vogelstein Adenoviral Plasmids Common shuttle and adenoviral plasmids. Want an adenoviral... Venus tag; See plasmid 62622 for plasmid without Venus Wang Adenoviral Kits Plasmids containing modular...Adenovirus Guide Virus Biosafety Lentiviral Plasmids AAV Plasmids Viral Vectors 101 eBook Adenoviruses are...incorporates sequences from both plasmids. This DNA molecule can then be transfected into mammalian packaging ...Strain for recombination between shuttle plasmid and adenoviral plasmid Vogelstein 16399 AdEasier-1 BJ5183 ...

...Collections Antibody Plasmid Collection Antibody Plasmid Collection Addgene distributes ready-to-use recombinant...Recombinant mAb Collection . Once the plasmids are transiently transfected into mammalian tissue culture cells... table highlights plasmid collections that can be used to create and express plasmid-based antibodies....Apr 8. Markus Seeger Antibody Plasmids The table below highlights plasmids that feature expression of antibodies...) ID Plasmid Description Gene/Insert PI Return to top Do you have suggestions for other plasmids that ...collection of plasmid-based antibodies, including monoclonal antibodies, scFvs, and nanobodies. Plasmid... Plasmid Collections...

... CRISPR Qi Lab CRISPR Plasmids Qi Labs CRISPR Plasmids Available from... genome. 1. Two-plasmid CRISPRi system for bacterial gene knockdown The first plasmid ( pdCas9_bacteria... both plasmids in bacteria could cause up to 300-fold repress on targeted genes. 2. Two-plasmid CRISPRi...fluorescent gating of transiently transfected cells. Co-expression of both plasmids in HEK293 cells could cause... 23849981 . Individual plasmids can be ordered via the links below: ID Plasmid Description 44246 pdCas9...fusion expression plasmid 44248 pgRNA-humanized A customizable gRNA expression plasmid for use with either...bacterial Cas9 expression plasmid 44250 pwtCas9-bacteria A wild-type Cas9 expression plasmid for use editing bacterial...

...Institute for Cell Science Plasmid Collection Allen Institute for Cell Science Plasmid Collection You May Also...full description of how to use these plasmids, check out Plasmids for Endogenous Gene Tagging in Human...Allen Institute for Cell Science has deposited plasmids to generate fluorescently tagged proteins of key...Fluorescent Protein Guide Subcellular Localization Plasmids Protocols The Allen Institute for Cell Science...induced pluripotent stem cell (hiPSC) lines. The plasmids used to generate these lines have been deposited...distribution to the research community. With these plasmids researchers will be able to generate fluorescently...Human Cells on the Addgene Blog. Plasmids for Endogenous Gene Tagging in Human Cells Find additional cell...

...publications associated with Addgene materials. Plasmids ID Plasmid Description PI Viral Preps Addgene distributes...research. Browse the BRAIN Initiative collection of plasmids, viral preps, and antibodies here. Collections...dissemination to the research community. Browse plasmids , viral preps , and antibodies created with support...distributes ready-to-use viral preps made from plasmids in our collection! AAV are distributed as high-quality...pAAV-CAG-FLEX-Archon1-KGC-EGFP-ER2 AAV production plasmid encoding for Archon1 fluorescent voltage reporter...IRES EGFP Chemogenetics Cre-dependent expression plasmid Scott Sternson 119741-AAV9 AAV SYN flex PSAM4 GlyR...IRES EGFP Chemogenetics Cre-dependent expression plasmid Scott Sternson 119742-AAV5 AAV SYN PSAM4 GlyR IRES...

...for select plasmids are now available through Addgene, learn more here . Browse Viral Plasmids by Type Visit... to find plasmids that can be used to engineer various types of viruses. Lentivirus Plasmids | Guide Lentiviruses...transgene expression. Overview of Viral Plasmids While plasmids are typically used to encode, propagate...organism. Figure 1. Using viral plasmids to generate virus. A) Viral plasmids have been engineered such that...genomes have been adapted into a plasmid-based technology, such that plasmids can be used to create viruses...researchers can introduce plasmids to a host to generate virus. Furthermore, these plasmids can be modified to...are usually divided among many plasmids, such that all of these plasmids must be present for a viral particle...

...in a new window) . Transfections are performed using the transfer plasmid, a plasmid encoding Rep and serotype-specific... coli . In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits. Resources... serotype-specific Cap genes, and a plasmid encoding adenoviral helper sequences. AAV preparations are...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...off-line and discarded. Confirmation of Transfer Plasmid Addgene uses a rigorous barcode matching system...microbial contamination. Purity When possible, all plasmids used for viral production are propagated in the...experiment using our virus protocols . Browse our plasmid collection of popular lentiviral vectors ....