...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 ...

...separate transfer, envelope, and packaging components of the virus onto different vectors. The transfer vector...Many lentiviral transfer plasmids are based on the HIV-1 virus. For safety reasons, transfer plasmids are...generation transfer plasmid cannot be used with a 3rd generation packaging system. What is the difference between...packaging; however, they are different viruses and thus use slightly different isoforms of these packaging...genome. Many lentiviral transfer plasmids do not have selectable markers conferring resistance to an antibiotic...Addgene's Plasmid Reference and Protocol Guide . How do I clone my insert into a transfer vector with no ... from the transfer vector during the viral production stage. Many of the lentiviral transfer vectors that...

... 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...

...Biosafety Guide Parts of an AAV Transfer Plasmid Video Web References Vector Biolabs: Introduction to ...adeno-associated viral vectors for enhanced gene transfer. Kwon I, Schaffer DV. Pharm Res. 2008 Mar;25(3):489-99....2, and 5 display differential efficiency and cell tropism after delivery to different regions of the central...Integration Serotypes Pseudotyping AAV Variants References AAV Components The small (4.8 kb) ssDNA AAV genome...VP3 - capsid proteins). When constructing an AAV transfer plasmid, the transgene is placed between the two... (E4, E2a and VA) mediate AAV replication. The transfer plasmid, Rep/Cap, and the helper plasmid are transfected...in optogenetics experiments. These viruses are preferred over lentiviruses because they remain primarily...

...pseudotype γ-retrovirus using different envelopes to modify tropism. Briefly, different envelope plasmids can ...engineered γ-retroviruses differ from lentiviruses? There are a few important differences between engineered ...packaging; however, they are different viruses and thus use slightly different isoforms of these packaging...technologies? No, the different “generations” associated with lentiviral transfer plasmids are determined... followimg components are needed: γ-Retroviral transfer plasmid encoding a transgene, sgRNA, or shRNA ...sequences, which facilitate integration of the transfer plasmid sequences into the host genome. Typically... the host genome upon viral transduction. Most transfer plasmids available from Addgene were derived from...

...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...

...pAdTrack Class of transfer vectors that contain IRES-GFP pShuttle Class of transfer vectors that do not...consists of two types of plasmids: shuttle (or transfer) vectors and adenoviral vectors. Find vectors ...Vogelstein designed multiple shuttle vectors for different purposes. The pAdTrack series contains an IRES-GFP...about Adenoviral Vectors Are adenovirus and AAV different? Yes! AAV is a small single-stranded DNA parvovirus...packaging plasmid or 911E4 packaging cell line Transfer Vocabulary Term Definition ITR Inverted terminal...

...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...