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Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

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We narrowed to 824 results for: EED

Showing: 761 - 780 of 824 results

  1. Plan Your Experiment

    Type
    Guide
    ...population need to be identical (clonal) or is a mixed population okay? Does the edit need to be homozygous...interference Needs good primer design Western blot Knockouts Activation/interference Needs a good antibody...other organisms. As with most experiments, you first need to have a biological question in mind. Then you ...will help you narrow down which components you’ll need for a given experiment. In the table below, we highlight...dCas9 to be fused to transcriptional activators that need to be within a given range of the transcription ...HEK293) cells. For more difficult cell types, you may need to look into other options. Browse Addgene's collection...cells are not transcribing DNA to mRNA, nor do they need to translate mRNA to protein. Instead, you directly...

  3. Antibody Guide

    Type
    Guide
    ...process may need to be optimized for individual assays. Clear cell debris by sedimentation. Proceed with IP..., an antimicrobial agent, can block some amines needed for conjugation reactions and should be removed...chosen. Signal Amplification The amount of signal needed from a specific antibody can vary depending on ...category, including an outline of the basic steps needed for each application, important controls, and any...samples to make proteins available. Tissue samples may need additional processing steps, such as sonication....signal levels. In ChIPseq, an input control will need to be collected before immunoprecipitation. Antibody...from. Then consider the following questions: Do you need polyclonal or monoclonal antibodies? Will you be...

  4. Adenovirus Guide

    Type
    Guide
    ...therefore needs to be supplied through other means. In order to produce rAdV vectors, E1 needs to be expressed...especially when needing transgenes to express for a shorter period of time or when needing to package larger...starts and primarily encode for structural proteins needed to assemble icosahedral capsids and build new virions...acronyms, so if you're new to viral vectors or simply need a refresher, we've included a glossary at the end... In order to produce rAdV vectors, you generally need two plasmids: Shuttle plasmid (also known as transfer...produced in high titers for their use in experiments. Needle sticks and ripped gloves are common points of entry...system for generating recombinant adenoviruses . Proceedings of the National Academy of Sciences of the United...

  5. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...transport relies on kinesin. Different rates of speed for various forms of retrograde and anterograde ...

  6. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ... easier for you to find and use the plasmids you need. You can find all the hot plasmids from 2017 below...

  7. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...acronyms, so if you're new to viral vectors or simply need a refresher, we've included a glossary at the end...In order to produce gamma-retroviral vectors, you need three plasmids (Figure 2): Transfer plasmid — contains...stably express gag-pol and/or env , eliminating the need to deliver these genes on separate plasmids ( in...with contact to mucous membranes or broken skin. Needle sticks and ripped gloves are common points of entry...clinical-grade gamma-retroviral vector purification by high-speed centrifugation for CAR T cell manufacturing. Molecular...specificity of infection for CD4-expressing cells. Proceedings of the National Academy of Sciences , 94 (16)...the beginning of R, has a 5' cap structure, and proceeds through U5 and the rest of the provirus. Some ...

  8. Cloning

    Type
    Guide
    ... there is an ideal approach. This may be due to speed, cost, availability of starting materials or just...compatible ends together. TOPO® cloning thus does not need restriction enzymes or an exogenous ligase providing... of PCR products into donor plasmids without the need for restriction enzyme cloning. For more information...of DNA in the chosen orientation, and without the need for any unwanted sequence at the junctions (such... a seamless manner. To accomplish this, you just need to introduce into the yeast the two (or more) fragments...would like to fuse. A major disadvantage is that you need to be set up to grow, transform and purify DNA from...

  9. CRISPR Guide

    Type
    Guide
    ...requirements for a CRISPR knockout assay are simple. You’ll need a Cas enzyme and a gRNA specific to the gene of ...Cas9-gRNA complex binds a putative DNA target, the seed sequence (8–10 bases at the 3′ end of the gRNA targeting...) will begin to anneal to the target DNA. If the seed and target DNA sequences match, the gRNA will continue...Mismatches between the target sequence in the 3′ seed sequence inhibit target cleavage, while mismatches...NHEJ directly ligates the break ends without the need for a homologous template. Both NHEJ and HDR have... the depositing lab. After amplification, you’ll need to use next-generation sequencing (NGS) to verify...matches that of the original library. Next, you’ll need to generate lentivirus containing the entire library...

  10. Lentiviral Vector Guide

    Type
    Guide
    ...acronyms, so if you're new to viral vectors or simply need a refresher, we've included a glossary at the end...table . In order to produce lentiviral vectors, you need three (or four, for third generation) plasmids: ...heterologous promoter (often CMV or RSV), eliminating the need for transactivation by Tat. Most third-generation...Biology Reference . Production After cloning, you will need to produce the lentiviral vectors from your plasmids...with contact to mucous membranes or broken skin. Needle sticks and ripped gloves are common points of entry...definition, at the beginning of R, is capped, and proceeds through U5 and the rest of the provirus. Third-generation...

  11. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...acronyms, so if you're new to viral vectors or simply need a refresher, we've included a glossary at the end...order to produce rAAV vectors, three plasmids are needed: Transfer plasmid (also known as cis plasmid) —.... The cell line used for rAAV vector production needs to express the adenovirus E1 gene, which is crucial...especially if any additional regulatory elements are needed. One strategy to overcome this obstacle, has been...with contact to mucous membranes or broken skin. Needle sticks and ripped gloves are common points of entry...

  12. Chemogenetics Guide

    Type
    Guide
    ...targets for development into chemogenetic tools. Need a refresher on G-protein coupled receptors? Read...back-metabolism of CNO to clozapine, demonstrating the need for alternative DREADD ligands. Compound 21, Deschloroclozapine...while PSAMs have more than one domain, both of which need to be expressed. However, neuronal control through...experiment, an AAV-encoded chemogenetic plasmid may need to be targeted to specific tissues, cell types or...

  13. Molecular Biology Reference

    Type
    Guide
    ...the genetic code and provides all the information needed for cells to make proteins and other molecules ...sequencing. To understand Sanger sequencing, we first need to understand DNA replication. DNA is a double helix...pieces of a 2,000 piece puzzle. A scientist would need to run a ton of Sanger sequencing reactions on different...Sequence FLAG DYKDDDDK HA YPYDVPDYA His HHHHHH Myc EQKLISEEDL V5 GKPIPNPLLGLDST Xpress DLDDDDK or DLYDDDDK ...

  14. Addgene's eBook Collection

    Type
    Guide
    ...When you're not sure where to start or need a handy reference, Addgene's eBooks are here to help!...Collection When you're not sure where to start or need a handy reference, Addgene's eBooks are here to ...101 blog series, this eBook covers everything you need to know about plasmids, from parts, nomenclature...

  15. Optogenetics Guide

    Type
    Guide
    ...Channelrhodopsin from Volvox carteri 570 Chronos High-speed, light-sensitive channel from Stigeoclonium helveticum...When designing your optogenetics experiment, you’ll need to pick both an opsin and a delivery system. Here...experimental design will determine whether you’ll need short or long periods of neuronal activation/inactivation...

  16. Guide to Using Pooled Libraries

    Type
    Guide
    ...amplified before you conduct your screen. If you need to amplify the library, please refer to the depositor...adequately represented in the population, you’ll need to infect many more cells than the number of plasmids...

  18. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...2022 Workflow Timeline Day 1: Seed cells Day 2: Transfect cells Day 3-6: Feed cells Day 7: Harvest antibody...fresh solutions after one month. BCD Feed 500 mL BalanCD HEK293 Feed 20 mL 200 mM Glutagro Store at 4 °C... °C. Procedure Section 1: Seeding cells The day prior to transfecting, seed a 108 mL culture of HEK293...post-transfection, supplement the flask with 4% BCD Feed. Pro-Tip The feed can be repeated up to 4 times for a total...Example feeding schedule: Thursday: Transfect cells. Friday (24 h post-transfection): Add 4% BCD Feed. Monday...to 3.75 mM, add 4% BCD Feed. Tuesday (120 h post-transfection): Add 4% BCD Feed. Wednesday (144 h post-transfection...purified for use in a variety of applications. Sharing speeds science. We believe that sharing the full details...

  19. Centrifugation

    Type
    Protocol
    ...control panel where you can set the time and speed needed for your experiment. Procedure An example of...centrifuge needed. This decision should be based on the size of tubes you are using and the speed at which...according to speed and time (and temperature, if applicable) listed in your protocol. Pro-Tip Spin speed is often...which is generated by spinning the sample at a high speed. Being able to separate solids from a liquid or ...accommodate different sized containers, spin at different speeds, or keep samples at specific temperatures. This...conical tubes. Depending on the centrifuge their speed ranges can vary, but are typically lower than some... about bench height. They often spin at similar speeds to their smaller cousins, but can hold much larger...

  20. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...QuickSeal tube spacers 16 ga needle 18 ga needle 10 mL syringe 18 ga blunt edge needles, Hamilton 1X PBS pH 7.4...Figure 2) with an 18 ga needle. Place the first microcentrifuge tube under the needle’s opening to collect ...interface with an 18 ga needle attached to a 10 mL syringe. The bevel of the needle should be up, facing ...a 10 mL syringe and a blunt edge 18 ga Hamilton needle, taking care to avoid bubbles (Figure 1). 8 mL ... 90 min in a T70i rotor at 10 °C. Pro-Tip If you need more time, you can alternatively centrifuge for ...Puncture the top of the QuickSeal tube with a 16 ga needle and start collecting 0.5 mL to 1 mL fractions per...AAV-containing Iodixanol solution will flow from the needle set at the 40–60% interface. Make sure that the...
Showing: 761 - 780 of 824 results