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TypeGuide...Packaging plasmid — contains packaging genes gag and pol Envelope plasmid — contains packaging gene env...recombination-deficient bacteria strains, such as NEB Stable cells. These strains reduce the frequency of homologous...expression in a host and low immunogenicity. This guide contains many viral vector-specific terms and acronyms,... into a host genome. This single strand of RNA contains packaging genes and the long-terminal repeats ... three plasmids (Figure 2): Transfer plasmid — contains transgene, sgRNA, or shRNA of interest flanked... greater yield of DNA. However, if the plasmid contains a Gateway cassette containing the ccdB gene, a... just a few days. Another variant, Phoenix-gp, contains only gag-pol and enables additional flexibility...
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Promoters
TypeGuide...factors, and other accessory proteins (such as activators and structural proteins) make up the transcription...distal promoter also contains transcription factor binding sites, but mostly contains regulatory elements...Constitutive Insect Strong promoter from Drosophila actin 5c gene Gal4/UAS Specific Insect Requires UAS...transcription factors. Transcription factors are proteins which recruit RNA polymerase and control and regulate...sequences, including enhancers, boundary elements, insulators, and silencers. Bacterial Promoters Promoters... can be either negative or positive. Repressor proteins bind to negative-repressible operons and prevent...region is located closest to the start codon and contains the RNA polymerase binding site, TATA box, and... -
Molecular Cloning Techniques
TypeGuide...amplify and manipulate genes of interest and then insert them into plasmids for replication and protein ...molecular cloning is the "classic" cloning method, and remains one of the most popular today. Restriction enzymes...identify compatible restriction enzymes that cut your insert or backbone at only the desired location for your...Restriction enzyme cloning of your gene of interest (insert) into a plasmid backbone. Created with BioRender.com... be cloned into a Gateway donor plasmid, which contains compatible attP sites via a proprietary BP clonase...any compatible Gateway destination vector, which contains attR sites, via LR clonase enzymes. Thus, you ...completed via Gateway cloning. Addgene's collection contains thousands of Gateway donor , Gateway entry , and... -
Plan Your Experiment
TypeGuide...small tag insertions, by copying from a DNA donor template Cas9 or Cas9 fusions with proteins that promote...including point mutations, base changes, and large insertions or deletions. For more information on the wide...our CRISPR Guide . There are many different Cas proteins available, and researchers can adapt these experiments...Cas9 nickase fusions with different base editing proteins) Many base editors fused to Cas9 exert their activity...sequence Base Edit Prime Edit Generate targeted insertions, deletions, and point mutations using a prime... In many cases, you can synthesize, anneal, and insert the targeting oligos into plasmids containing the...can generate stable cell lines if your plasmid contains a selection marker. Plasmid delivery is versatile... -
Modular Cloning Guide
TypeGuide...successive assembly steps. Before beginning, scientists insert fragments of DNA containing basic parts (promoters...optimize secretion and cell surface display of proteins of interest. Yeast Secrete and Detect Yeast Expression...cerevisiae . MoClo Baculo Toolkit Yeast Expression, Insect Expression Chen Davidovich 41 plasmids to construct...terminators, protein purification tags and fluorescence proteins for use in E. coli for a variety of applications... and Cultivarium Tools 189 plasmids, including origins of replication and selection markers for a variety...methylation-switching. 4G-Cloning Plasmid Kit Bacterial Expression, Insect Expression, Mammalian Expression Stephan Gruber...Zhao Plasmids to assemble custom-made RNA-binding proteins based on the Pumilio and FBF (PUF) homology domain... -
Addgene's eBook Collection
TypeGuide...Select Fluorescent Proteins 101 August 2017 | First Edition About Fluorescent Proteins 101 August 2017...Second Edition About This guide to viral vectors contains an introduction to each vector and its applications...2024 | First Edition About Our newest eBook contains practical introductions and helpful tips for antibody-based... -
Educational Resources
TypeGuide...Plasmids 101 CRISPR 101 Viral Vectors 101 Fluorescent Proteins 101 Science Career Guide Videos Watch our collection...Topics include: Genome Engineering Fluorescent Proteins CRISPR Viral Vectors Plasmid Technology Career...Restriction Digests Guides Browse our in-depth instructional content on subjects such as: CRISPR Chemogenetics... -
Science Guides
TypeGuide...system, antibodies are proteins capable of highly specific binding to other proteins. Antibody-based applications...can be used to capture, quantify, and identify proteins of interest. Read More Chemogenetics Chemogenetics... -
Guide to Using Pooled Libraries
TypeGuide... each plasmid contains a unique cDNA. In shRNA or gRNA libraries, each plasmid contains a unique gene ... -
Sequencing Primers
TypeGuide...GCTTGGATACACGCCGC MMLV sequence, for inserts in pBMN retroviral vector MMLV sequence, for inserts in pBMN retroviral vector...