...Educational Resources Molecular Biology Reference Molecular Biology Reference Jump To: Molecular Biology Basics...contain a reporter gene (for example, luciferase or GFP) that offers a read-out of the activity of the genetic... C EDQVDPRLIDGK S Tag KETAAAKFERQHMDS SB1 PRPSNKRLQQ Webpage and Blog References Addgene's blog , including...Addgene's molecular biology reference guide, including information on molecular genetics, plasmids, sequencing...verification Resources Genetic Code Webpage and Blog References Protocols Origins of Molecular Genetics The concept... Joshua Lederberg coined the term plasmid, in reference to any extrachromosomal heritable determinant....bacterium to another by a type of horizontal gene transfer (conjugation), usually providing a benefit to ...

...packaging genes. However, they are different viruses and require different isoforms of these packaging components...second-generation transfer plasmids as well. Third-generation plasmids include (Figure 4): Transfer plasmid — ...vectors separate transfer, envelope, and packaging components of the virus between different plasmids. The...activate the promoter on the transfer plasmid. Many of the lentiviral transfer plasmids that have been deposited...genes. From an experimental standpoint, the main difference between lentiviruses and gamma-retroviruses is...three (or four, for third generation) plasmids: Transfer plasmid — contains transgene, sgRNA, or shRNA ...systems, due to a wide range of infectivity for different cell types (known as tropism). Lentiviral vectors...

... dates. Buffers Buffers are a concern for conjugating and signaling molecules, as some buffer components...heavy chain constant region” or Fc. Different isotypes play different roles in the immune response, and ...removed from the buffer before the reaction. However, it can be added back to buffers after the conjugation...of colors activated by different wavelengths, allowing for easy differentiation between multiple antibodies... In some cases, it may be preferable to buy small aliquots of many different antibodies and do batch validation...an antigen, while the specific binding site is referred to as an epitope. Production Antibodies are typically...typically produced in one of three different methods: in animals, in cultured immune cells, or through...

... via a different plasmid. In order to produce rAAV vectors, three plasmids are needed: Transfer plasmid...s Molecular Biology Reference . Production The three production plasmids (transfer, packaging, and helper...2, and 5 display differential efficiency and cell tropism after delivery to different regions of the central...Integration AAV Types Common Uses Safety Resources and References Plasmid Elements Glossary Adeno-Associated Viruses...immune response. These features have made AAVs the preferred choice to be used as viral vectors for gene therapy...transgene, gRNA, or shRNA of interest into the transfer plasmid can be done with most standard cloning...restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited restriction sites or...

...three different plasmids containing the transfer, envelope, or packaging components. The transfer plasmid...packaging genes. However, they are different viruses and require different isoforms of these packaging components...type. From an experimental standpoint, the main difference between lentiviruses and gamma-retroviruses is...gamma-retroviral vectors, you need three plasmids (Figure 2): Transfer plasmid — contains transgene, sgRNA, or shRNA ... gene, due to a wide range of infectivity for different cell types (known as tropism). Figure 2: Gamma-retroviral...transgene, gRNA, or shRNA of interest into the transfer plasmid can be done with most standard cloning...restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited restriction sites or...

... (A to G). Different serotypes exhibit varying tissue tropisms, meaning they preferentially infect certain...from different species. Used to alter infectivity and tropism. pShuttle Class of shuttle/transfer vectors...Adenoviral Types Common Uses Safety Resources and References Plasmid Elements Glossary Adenoviruses (AdV) .... These genes are now supplied in trans , via different plasmids or the packaging cell line, which not...need two plasmids: Shuttle plasmid (also known as transfer plasmid) — containing the transgene of interest...recombination between the transfected adenoviral transfer plasmid and the E1 region integrated in the packaging...vectors Third-generation rAdV vectors are also referred to as "gutless" or high capacity AdV (HCAdV) vectors...

...Alternatives Resources Plan Your Experiment Web References PAM Sequences Glossary Publications CRISPR Overview...techniques . Collectively, these techniques are referred to as CRISPR ( C lustered R egularly I nterspaced...measure of fidelity, so sometimes they will be referred to as increased fidelity enzymes. hfCas9s were...pyrimidine-to-pyrimidine) Base transversions - edit to different classification of nucleotide base (purine-to-pyrimidine...opposite strands Prime editing and base editing both offer high efficiency and low indel formation compared...genetic changes, including moving entire genes to different locations, by combining other gene editing mechanisms...DNA-targeting of CRISPR-Cas systems. You may also see them referred to as INTEGRATE ( I nsertion of T ransposable ...

...light-emitting luciferase and a light-sensing optogenetic element. When the luciferase substrate (luciferin) is ...efficiencies. Depending on the type of luciferase used, the luciferin substrate may be the native coelenterazine...activity in neurons LMO Luciferase Opsin Ligand Effect Outcome (in neurons) Reference LMO3 sbGLuc VChR1 CTZ...RASSLs DREADDs PSAMs LMOs Planning Your Experiment References Chemogenetic tools are actuators for specific... of chemogenetic receptors. There are several different types of DREADDs that can be broadly classified...inhibitory KORD DREADD was developed to respond to a different ligand, Salvinorin B (SALB). Scientists have shown...Receptor Effector Ligand Effect Outcome (in neurons) Reference Rq(R165L) Human M3 muscarinic Arrestin-2/-3 CNO...

...Introduction Microbial Opsins Optical Switches References Introduction The field of optogenetics integrates...naturally occurring microbial opsins that respond to different wavelengths of light, like blue or yellow light...light. These various opsins also initiate different electrochemical responses, such as nonspecific cation... blue light activated cation channel. ChR2 is preferred over ChR1 because ChR2 has higher conductance ...optimization, and chimeric fusion of domains from two different ChRs. These ChR variants still function as light-gated...Color of activation light. There are a variety of different activation wavelengths, ranging from blue to yellow...rendering the experimental procedure less invasive. Different activation wavelengths also make it possible to...

... Plasmid Reference Molecular Biology Reference Sequencing Primers Sequencing Primers...convenience. For reference information, please consult Addgene's Molecular Biology Reference Page . All listed...AGTCAAGTAACAACCGCGA 3' end of luciferase, forward primer LucNrev CCTTATGCAGTTGCTCTCC 5' end of luciferase, reverse primer...sequences listed on the left are provided for your reference. Addgene does not distribute primers. For sequencing... primer LucNrev CCTTATGCAGTTGCTCTCC 5' end of luciferase, reverse primer M13 Reverse CAGGAAACAGCTATGAC...GGGCTGGCAAGCCACGTTTGGTG 3' end of glutathione-S-transferase, forward primer SP6 ATTTAGGTGACACTATAG SP6 promoter...GGGCTGGCAAGCCACGTTTGGTG 3' end of glutathione-S-transferase, forward primer pGP704-R AACAAGCCAGGGATGTAACG...

...Desired Genetic Manipulation Different genetic manipulations require different CRISPR components. Selecting...be present in a single lentiviral transfer vector or separate transfer vectors May contain reporter gene...experimental goal, you are ready to start navigating the different reagents that are available for your particular...particular cell type or organism without a specific preferred mutation Cas9 (or Cas9 nickase) Single (or dual...can make a limited set of mutations. Repress or Interfere (CRISPRi) Reduce expression of a particular gene...targeting promoter elements of target gene Many different activators exist, including the multi-plasmid ...transduction CRISPR elements are inserted into an AAV transfer vector and used to generate AAV particles (for...

... Plasmid Reference Molecular Biology Reference Promoters Promoters Definition A promoter is...Addgene's molecular biology reference material covering promoters, transcription and the transcription...mRNA transcript produced. The antisense strand is referred to as the template strand or non-coding strand...transcription of DNA into mRNA. Promoter binding is very different in bacteria compared to eukaryotes. In bacteria...promoter region. Each sigma factor recognizes different core promoter sequences. Operons Although bacterial...regulation, like operons. Operons are a cluster of different genes that are controlled by a single promoter... types of RNA polymerases that all transcribe different genes. RNA polymerase I transcribes genes encoding...

...you to do many different molecular biology techniques (such as fusing it with different tags, putting ...Addgene's reference guide on molecular cloning techniques, including information on seven commonly used... Molecular Biology Reference Choosing a Molecular Cloning Technique Molecular Cloning...availability of starting materials or just personal preference. Check out our blog on choosing the right cloning...Gateway® destination plasmids have been made with different promoters, tags and fluorescent proteins. Thus... variety of promoters and into backbones with different selection cassettes). Although gateway cloning...

...pooled libraries are single preparations of many different plasmids. Plasmids within a given library have...the same backbone, but they express or target different genes. Some libraries cover the majority of the... library from the Moffat lab has over 175,000 different gRNA-containing plasmids). Amplifying and Using...screen. If you need to amplify the library, please refer to the depositor’s protocol for the best results...conducting pooled library screens. Addgene does not offer next-generation sequencing services. For an NGS ...Genome-Scale, Knockout Screening Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens...

...Pooled libraries are single preparations of many different plasmids. Plasmids within a given library have...the same backbone, but they express or target different genes. Some libraries cover the majority of the...Find educational guides to learn more about the different types of viruses that are commonly used in research...

...you're not sure where to start or need a handy reference, Addgene's eBooks are here to help!...you're not sure where to start or need a handy reference, Addgene's eBooks are here to help! 0 of 0 Selected...and management, resources for identifying your transferable skills, and tips on the job application process...