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...Additional Cloning Information Cloning Protocols Single guide cloning: Zhang Lab General Cloning Protocol...provided as empty backbones that utilize Golden Gate cloning to insert the desired guide RNA sequence by digestion... annealed and phosphorylated oligonucleotides containing the guide target sequence: Up to two guides can...Protocol (PDF, 237 KB) Dual guide cloning: Brzezinski Lab Dual Guide Protocol (PDF, 990 KB) Plasmid and Sequence...

...Proteins FREQ-Seq Kit FX Cloning Kit GreenGate Cloning System (Plant) MoClo Modular Cloning Modular PUF Assembly...Plasmids by Category Cloning and Genomic Tools Browse plasmids related to cloning and genomic modification...Pamela Silver Lei Stanley Qi Jeff Tabor Jan-Willem Veening Christopher Voigt Clifford Wang Ron Weiss Synthetic... Networks and Gene Regulation Browse plasmids containing both naturally occurring and synthetic regulatory...

...Packaging Service page . Pooled CRISPR Libraries for Screening Use pooled CRISPR libraries to screen for genes...backbone lentiGuide-Puro targeting 19,114 genes and containing 76,441 unique sgRNAs along with 1000 non-targeting...backbone lentiCRISPRv2 targeting 19,114 genes and containing 76,441 unique sgRNAs along with 1000 non-targeting... XPR_502 (P65 HSF) targeting 18,885 genes and containing 56,762 unique sgRNAs. This backbone does not ...backbone pXPR_050 inhibiting 18,901 genes and containing 57,050 unique sgRNAs along with 500 non-targeting...backbone pXPR_050 inhibiting 18,899 genes and containing 57,011 unique sgRNAs along with 500 non-targeting...Mouse sgRNA library in backbone lentiGuide-Puro containing 78,637 unique sgRNAs targeting 19,674 genes along...

...electrophoresis (PAGE) followed by silver staining or SYPRO Ruby staining and the molecular weight and relative...after negative staining. Empty vector particles can be identified after negative staining and appear darker...purity of the AAV preparation. Figure 1: Silver staining of purified and non-purified AAV subjected to ...Microscopy The ratio of empty to full (i.e., genome containing) AAV particles within representative vector preparations...Trono lab . Cell culture medium (OptiPro SFM) containing lentivirus is first cleared by low-speed centrifugation...

...proteins and empty backbones for expression and cloning of fusion proteins, along with key parameters to...506 517 93 5.7 10 min Monomer pCS2+mNeonGreen-C Cloning Vector - Mammalian Expression Return to top Yellow...Zebrafish/Xenopus/Worm/Sea urchin pET mCitrine LIC cloning vector (u-mCitrine) - Bacterial Expression mCitrine-pBAD...mOrange 548 562 49 6.5 2.5 h Monomer pET mOrange LIC cloning vector (u-mOrange) - Bacterial Expression pCS2+...pmScarlet_C1 - Mammalian Expression pCS2+mScarlet-C Cloning Vector - Mammalian Expression mScarlet-I 569 593...587 610 16 4.5 15 min Monomer pcDNA3 mCherry LIC cloning vector (6B) - Mammalian Expression pLV-mCherry ...Zebrafish/Xenopus/Worm/Sea urchin pET mCherry LIC cloning vector (u-mCherry) - Bacterial Expression pFA6a-link-yomCherry-SpHis5...

...well as by our trusted partner labs. Learn More Cloning and Expressing Antibodies Antibody Plasmids Antibodies...synthetically produced recombinant antibodies, created by cloning antibody components into plasmids and expressing...and their applications in our Antibody Guide ! Cloning and Expressing Antibodies The following table highlights...mammalian expression vector A tool kit for rapid cloning and expression of recombinant antibodies. Sci Rep... Rep. 2014 Jul 30;4:5885. Andrew Beavil Cloning and mammalian expression of mouse and human IgG monoclonals...and secreted proteins suitable for wholemount immunostaining. Biochem Biophys Res Commun. 2015 Jan 2;456...

...targets adenenine, NG targets thymine, and NN targets guanine (though NN can also bind adenenine with lower...approximately every 35bp. Researchers are still determining the importance of context for each TAL effector...be used with the recently developed Golden Gate cloning method to assemble “multiple DNA fragments in an...to hone these tools and the researchers who are learning to wield them. Learn more about Addgene’s TAL ...

... generally called pTet) have been designed by combining tet O sequences with elements like the H1 RNA ...TetO" or "Tet-O" in their name or description, meaning they contain a TRE or tet O and can serve as either...tetracycline or its derivatives which can result in unintended induction. Check the manufacturer’s details to...Backbones Find backbones or constructs to use for cloning to control expression of your gene of interest....EZ-Tet-pLKO-Puro Lentiviral Tet-On plasmid for easier cloning and design of inducible shRNA expression, with ...Sleeping Beauty transposon system with luciferase in cloning site. See article ( Kowarz et al., 2015 ) for additional... for inducible expression; insert with Gateway cloning and puromycin selection rtTA-Advanced Tight TRE...

...assembly, and the remaining 5 plasmids include Cre and FLP expression vectors, a general cloning plasmid, and...plasmid-system consists of two components: A vector containing Cas9 and a validated gRNA, based on the Zhang... also provided their protocol for homology arm cloning: Mendenhall & Myers FETCh-seq Protocol 134.7 KB...system depends on MMEJ (microhomology-mediated end-joining), an alternative DSB repair pathway, instead of...were created using CRISPR and the donor plasmids containing homology arms and EGFP are available at Addgene...

...available at Addgene. The collection can be used for screening transcription factors for cellular programming... The individual clones, or the pooled library containing the entire pooled collection, can be purchased.... This collection and library can be used for screening transcription factors for applications such as...window) . TF ORF barcode PCR for NGS QC and bulk screening (PDF, 85 KB) TF ORF barcode dial out PCR for 10X...bp sequence, to simplify identification during screening. Library ID PI Description MORF library 137000...

...kinase that anchors a signaling network for transitioning between anabolic and catabolic states. Ras Ras...from the Boehm , Hahn , Meyerson , and Root Labs containing wild-type and mutant alleles described in the... Set of vectors from the Sabatini & Wood Labs containing cDNAs for studying oncogenic signaling pathways...from the Esposito Lab for use with the Gateway® Cloning Platform (Life Technologies™) to permit construction... Tackling Cancers’ Drug Resistance with a New Screening Kit (David Sabatini & Kris Wood) National Cancer...

...double-stranded siRNA and degrades one of the strands. The remaining single-stranded siRNA then binds the target sequence...Plasmids Browse our selection of empty vectors for cloning in shRNAs. Most plasmids are in a lentiviral backbone...Tet) expression are available. To find plasmids containing other RNAi components, like argonaute, please... Additional Resources Addgene Resources pLKO.1 cloning vector protocol Other Addgene viral protocols Lentiviral.... Wiznerowicz, M., Szulc, J., Trono, D. (2006). Tuning silence: conditional systems for RNA interference...

...base editors — cytosine base editors (CBEs) and adenine base editors (ABEs) — can be used to generate single...or a G to A on the opposite strand). Likewise, adenine base editors have been engineered to convert adenosine...the cell, creating an A to G (or T to C) change. Adenine DNA deaminases do not exist in nature, but have...evolution of the Escherichia coli TadA, a tRNA adenine deaminase. Like cytosine base editors, the evolved...domain is fused to a Cas9 protein to create the adenine base editor. Both types of base editors are available...

...purification tags compatible with Golden Gate modular cloning (MoClo) for use in E. coli . Addgene Blog E. Coli...together and reconstruct the full FP. Find plasmids containing FPs and split FPs that can be fused to your proteins...Lactose/IPTG Streptococcus pneumoniae Jan-Willem Veening 188978 pREDawn-AmpR-MCS FixK2 Red light (660 nm...molecules in and around a bacterium. Plasmids containing easily measurable reporter genes (e.g., LacZ,...Bacterial One-Hybrid (B1H) System can be used for determining the DNA-binding specificity of proteins such ...Plasmids Collection for a selection of plasmids containing recombinases to induce targeted genetic inversion...

...that are amenable to genetics, high-throughput screening and non-invasive live imaging. A standard toolbox...Tools for Use in Zebrafish Looking for plasmids containing your zebrafish gene of interest? Search Addgene's...biotin labeling in zebrafish. pGCS Vector Kit - Hai-Ning Du Lab. A set of pCS2+ backbone-based Gateway destination... Amro Hamdoun Lab. A useful “starter kit” for screening fluorescent protein fusion expression in organisms...zebrafish as a model organism and to foster advanced training of young scientists in Latin America. The Zebrafish...

...antibodies. Recombinant antibodies are created by cloning antibody genes from specific hybridoma cells into... two types of antigen specificity: monoclonal, meaning many copies of one clone that are specific to a... a certain epitope, or polyclonal, meaning there are many clones that are specific to multiple epitopes...using permeabilization, antigen retrieval, and/or sectioning. Block to reduce non-specific binding of antibodies...panels used currently are in the mid-forties. When designing a multiplex assay, selection of the fluorophores... your target epitope in the assay type you are running. Sufficient validation data specific to your antibody...Validation of an antibody is a process separate from running appropriate controls in your assay. There are five...

...Receptor Binding Domain Deep Mutation Scanning Library Screening Jesse Bloom Library of Spike protein ...called SARS-CoV-2. In 2020, scientists worked at lightning speed to elucidate as much as they could about...-CoV-2 Spike (S) Ectodomain and RBD Libraries Screening Timothy Whitehead Libraries of Spike (S) Ectodomain... Domain Site-Saturation Mutagenesis Libraries Screening Jesse Bloom Tyler Starr Libraries of variant Spike...

...unique potential to continuously divide while maintaining an undifferentiated state. Stem cells are classified...Sox2, Klf4, and c-Myc from four separate plasmids Defining molecular cornerstones during fibroblast to iPS...Neurons Lentiviral Human Small molecules enable neurogenin 2 to efficiently convert human fibroblasts into...human fibroblasts using self-regulating and nonintegrating viral vectors. Stem Cell Reports. 2014 Nov ...

... Tackling Cancers’ Drug Resistance with a New Screening Kit Background mTOR Pathway Plasmids mTOR Gene... growth and proliferation. mTOR, a serine/threonine kinase, is found in two complexes, mTORC1...LST8 homolog LKB1 Also known as STK11; serine/threonine kinase 11 mTOR Mechanistic target of rapamycin... Tackling Cancers’ Drug Resistance with a New Screening Kit (from David Sabatini & Kris Wood) References...

...mutant to inhibit MMR PEmax — optimized PE2 enzyme containing RT optimized for human codons, additional nuclear...check out our prime editing blog post . Need help designing a pegRNA? Try the DeepPrime (Link opens in a new...of gRNA (e.g., pegRNA, epegRNA, nicking sgRNA), cloning enzyme, selectable marker, and whether the plasmid...Plasmid Expression System Promoter Guide RNA Type Cloning Enzyme Co-expressed Cas9 Selection PI 132777 pU6...