Skip to main content
Addgene
Open Menu

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

Information

In Category

We narrowed to 940 results for: Mpl;

Showing: 101 - 120 of 940 results

  1. CRISPR Guide

    Type
    Guide
    ...genomic edits by copying from a DNA donor template . The donor template contains the desired sequence and is...linear dsDNA templates work better for large edits or insertions. When designing the donor template, incorporating...experiments and must be amplified before use (Figure 8C), following the amplification protocol specified by...Cascade ( C RISPR- as sociated c omplex for a ntiviral de fense) complex to initiate DNA degradation. Cascade...Cascade C RISPR- as sociated c omplex for a ntiviral de fense; a complex of multiple Cas enzymes including... every genomic target. Due to its comparative simplicity and adaptability, CRISPR rapidly became the most... change the genomic target of the Cas enzyme by simply changing the target sequence present in the gRNA...

  2. Optogenetics Guide

    Type
    Guide
    ...Human codon optimization Example: Halo Increased photocurrent amplitude Examples: eNpHR, eNpHR2.0, eNpHR3.0...Increased light sensitivity Example: ArchT Increased photocurrent amplitude Examples: eArch3.0, eArchT3.0 Browse...enhancements include: Increased photocurrent amplitude Examples: ChR2(H134R), C1V1(t/t), ChIEF Increased ...) kinetics: Examples: ChETA, C1V1(t/t), ChrimsonR Red-shifted peak action spectra Examples: VChR1, C1V1...enhancements include: Increased photocurrent amplitude Examples: iChloC, SwiChRca, Phobos, Aurora Browse ...enhancements such as: Improved photocurrent amplitude Example: eMac3.0 Browse Leptosphaeria plasmids . Glossary... cells in order to understand their functions. Example schematic of an optogenetics procedure. A channelrhodopsin...

  3. Molecular Biology Reference

    Type
    Guide
    ... DNA, or templates, are spatially separated. These attached DNA templates are then amplified again, producing...producing ~1,000 copies of each template. Each template is then replicated using the modified bases and...plasmid using bacteria, which take up plasmids and amplify them during cell division. Because bacteria are... vary broadly in their functionality. In their simplest form, plasmids require a bacterial origin of replication... for the ability of the plasmid to be copied (amplified) by bacteria. Antibiotic Resistance Gene Allows...sequence used as an initiation point for PCR amplification or DNA sequencing of the plasmid. Primers can... DNA fragments. Cloning vectors tend to be very simple, often containing only a bacterial resistance gene...

  4. Guide to Using Pooled Libraries

    Type
    Guide
    ...see if it should be amplified before you conduct your screen. If you need to amplify the library, please...recommended to verify that the library is complete - an incomplete library may lead to false positives or...types of screen employ a selection method relevant to the phenotype being studied. Examples of published ... Pooled Library Amplification Protocol for Addgene's generalized library amplification protocol. Read ...Overview of what pooled libraries are; how to amplify and use a pooled library; and what types of screens...: Pooled Library Guide Introduction Library Amplification Types of Screens Resources An Introduction to...genes that contribute to a phenotype. At their simplest level, pooled libraries are single preparations...

  5. Molecular Cloning Techniques

    Type
    Guide
    ...discovery. With molecular cloning, scientists can amplify and manipulate genes of interest and then insert...overhang (blunt ends). Two pieces of DNA that have complementary overhangs, or which are both blunt-ended, can...restriction sites to any piece of DNA during PCR amplification, allowing for it to then be digested and ligated...a single recombination reaction, drastically simplifying the process and reducing the amount of time required.... To do this, your DNA fragment must first be amplified with specific Gateway attB1 and attB2 sites attached...experimental goal. Although Gateway cloning is a simple and efficient cloning method, Gateway vectors and...restriction cloning, once your project has been completed via Gateway cloning. Addgene's collection contains...

  6. Promoters

    Type
    Guide
    ... transcription is simpler than eukaryotic transcription, bacteria still have complex systems of gene regulation... including transcription and the transcription complex. Educational...produced. The antisense strand is referred to as the template strand or noncoding strand, as this is the strand... transcribed by the RNA polymerase. Figure 1: Simplified promoter region during transcription. Created...other hand, the process in eukaryotes is much more complex. Eukaryotes require a minimum of seven transcription...for transcription initiation. The -35 position, simply titled the -35 element, typically consists of the...transcription. The Lac operon in bacteria is an example of a negatively-controlled operon (Figure 3). A...

  7. Plan Your Experiment

    Type
    Guide
    ... donor template Cas9 or Cas9 fusions with proteins that promote HDR Requires a repair template Lower editing...edits using CRISPR/Cas9 in mammalian cells as an example, but many of these principles also apply to using... process of interest. Do you want to: Generate complete and permanent loss of gene expression or function...Remember that HDR also requires donor DNA to serve as template for the repair. Once you settle on your gRNA, ...Editing : pegRNAs must serve as both a guide and a template. In general, edits must be downstream (3’) of ...edit site yield higher editing efficiencies. The template sequences can also have a large impact on editing...perfect world, your gRNA sequence would be an exact complement to your target sequence with no homologous sites...

  8. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...as adenovirus or herpes simplex virus to replicate in the host cell and complete the lytic cycle. AAVs ... acronyms, so if you're new to viral vectors or simply need a refresher, we've included a glossary at ...transduction efficiency and alter tropism. For example, AAV2/5, targets neurons that are not efficiently...transduction efficiency and viral tropism. One common example is AAV-DJ, which contains a hybrid capsid derived...Several methods have been developed and successfully employed to engineer new rAAV capsid variants, such as ...retrograde (AAVrg) Directed evolution has also been employed to engineer an rAAV serotype variant able to display...blog post on AAVrg for more information. Self-complementary AAV (scAAV) One downside of the use of AAVs ...

  9. Modular Cloning Guide

    Type
    Guide
    ...restriction cut site, multiple modules with complementary overhangs can be digested and ligated in a single...vector creating a single transcriptional unit (for example, a promoter, a 5' UTR, a signal peptide, a protein-coding...Level 2 vectors permits the creation of even more complex constructs constrained only by the ability of E...overexpression, gene editing, functional genomics, and more complex metabolic engineering and synthetic biology, compatible... MoBiFC - Modular Bimolecular Fluorescence Complementation Toolkit Plant Expression Benjamin Field 50 ...plasmids for assembling bimolecular fluorescence complementation experiments to analyze protein-protein interactions...toolkit with both high cloning capacity and vector simplicity, compatible with other MoClo, GoldenBraid, and...

  10. Adenovirus Guide

    Type
    Guide
    ... acronyms, so if you're new to viral vectors or simply need a refresher, we've included a glossary at ...due to their well-established methods, relative simplicity of production, and higher titers. However, the...also resulted in a reduction of viral vector amplification and overall lower viral titers. Because of this...transduction efficiency, tropism and immunogenicity. For example, Ad5/35 and Ad5/26 combine the high transduction...packaging capacity. Large protein-coding genes and complex gene expression cassettes are often too big to .... K. (2011). Adenoviral vector immunity: its implications and circumvention strategies . Current Gene ...., Kinzler, K. W., & Vogelstein, B. (1998). A simplified system for generating recombinant adenoviruses...

  11. Chemogenetics Guide

    Type
    Guide
    ...any off-target effects. They are the most widely employed type of chemogenetic receptors. There are several..., and hM5Dq, however, hM3Dq is the most widely employed. Gi-DREADDs signal through the Gαi/o G-protein...allowed them to respond to their cognate PSEM. For example, PSAM Q79G,Q139G is activated by PSEM 22S , while...will determine the chemogenetic ligand used. For example, a study in rats using an hM4Di receptor might ...adrenergic receptor to engineer a receptor-effector complex for gene therapy. J Biol Chem. 276:1596-6. PMID...

  12. 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 ...hence the use of “retro”. Lentiviruses have more complex genomes, containing packaging genes and accessory...along with virus-specific accessory genes (for example, vif , vpr , vpu , and nef for HIV-1). The LTRs...Timms et al. (2018), and Wells et al. (2020) for examples). Pseudotyping Tropism dictates which types of...

  13. 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 ... — hence the use of “retro”. Retroviruses have simpler genomes, containing only the necessary packaging...transfection step of the viral packaging cell line. For example, Phoenix™ is a popular second-generation gamma-...

  14. Which Fluorescence Microscopy Technique is Best for Me?

    Type
    Blog Post
    ... dimensional sample (see Figure 1); similar to an MRI or CT scanner. This is accomplished by focusing ... to a single focal point. This is accomplished by using, for example, red light instead of blue to excite...between the glass coverslip and a sample in aqueous buffer it is completely reflected. This reflection occurs...water-like buffer the sample is immersed in. Although the excitation light is completely reflected, energy...my experiment? When I am contemplating which microscope to use for a new sample I always start by asking...asking two questions: 1) Is the sample dynamic or static? 2) Is the sample thin (< 15 μm) or thick? These...capture in these types of samples is their fast movements. Depending on the sample, a temporal resolution...

  15. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    ... multiplex PCR:  Single template PCR reaction - one template is amplified using several forward and ...PCR requires free nucleotides (dNTPs), template DNA to amplify from and unique single stranded DNA primers...reaction:  Denaturation- To amplify DNA, the two strands of the template DNA first have to be separated...End point PCR’s goal is to amplify a specific target of DNA. This amplicon can be used in molecular cloning...determine the concentration of the original template DNA in the sample.  Addgene currently uses ddPCR for AAV...Multiplex PCR, as the name implies, is a method in which multiple targets can be amplified in a single PCR experiment... reverse primer sets.  Multiple template PCR - multiple templates with different primer pairs that align...

  16. A Control for All Seasons

    Type
    Blog Post
    ...control is generally a sample or group that will have a desired response. For example, if you are studying...control is a sample or group that is not subjected to the experimental condition. In the example above, a ... wild-type and knockout samples. In this test, wild-type and knockout samples are treated with the identical...present in the wild-type samples and absent in the knockout.  If knockout samples are unavailable, try substituting...interactions between the sample and the secondary antibody. C, a label control treats the sample to the full protocol...endogenous antibodies naturally present in the sample, for example at sites of inflammation. This tends to be...all new samples, protocol changes, or when high background labeling occurs (Burry 2011). Complex experiments...

  17. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...in a PCR protocol that amplifies the entire plasmid template. The parent template is removed using a methylation-dependent...requires a template of high homology to the endogenous target, yet to facilitate the repair, the template should...mutagenized plasmids. PCR primers (green) amplify the plasmid template (blue), and introduce a restriction ... flank the targeted region. Upon PCR amplification, the template is removed by DpnI restriction digestion...are designed to be complementary, but each primer may extend beyond the complementary region as long as...efficiently amplified than larger constructs, but plasmids as large as ~6 kb can be amplified fairly easily...extension time to match the size of your template. Amplification of GC-rich plasmids is facilitated by the addition...

  18. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...with each sample. Successfully analyzed samples will display the following parameters: Sample - The unique...multiplex sample, ICE includes visual representations of all detected edit types in the sample. The visualization...indel.   For a complete guide to ICE analysis, including detailed explanation and examples, read the paper...easy. Simply upload your Sanger sequencing files and provide basic information such as your sample names...the ICE software has two analysis formats: sample by sample analysis, which can compare up to five editing...results and a list of the analyzed samples (see below).   If the sample run had no issues, the analysis ...perform more in-depth analyses on each sample by clicking on the sample name or on its corresponding bar graph...

  19. What’s in Your Tube?: A Quick Guide to Measuring DNA by Spectrophotometry

    Type
    Blog Post
    ... easy to measure sample concentration and purity using only a tiny bit of the sample itself — so easy ...instrument can calculate a sample’s nucleic acid concentration based on the sample’s absorption at 260 nm (...spectra of a mixed sample.  But, other molecules that may be present in the sample also absorb light...levels exceed 70% of the sample content (Loughrey and Matlock, 2016). So, even samples with “good” A260/280...nm, you may have phenol in your sample.  RNA contaminants DNA samples can also be contaminated with RNA.... To confirm RNA in your DNA sample, you will need to use a complementary assay, such as a fluorescence...especially problematic in low-concentration samples (samples with concentrations < 10 ng/μL), where absorbance...

  20. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...generation simpler, as described in an earlier post on generating CRISPR mouse models. For simple knockout... Single-stranded DNA repair templates Short single-stranded DNA templates have been shown to improve the...final ssDNA template. Figure 2: Overview of ivTRT to generate an ssDNA repair template. Although...ssDNA template and CRISPR machinery, Quadros et al. recommend ribonucleoprotein (RNP) complexes of Cas9...Easi-CRISPR. As in the knock-in example, you’ll design your repair template with 55-100 bp homology on each...plenty of Addgene plasmid options for templates - see the example at right using blue flame plasmid pmScarlet_C1.... It’s also made targeted insertions relatively simple in C. elegans and bacteria. But CRISPRing typical...
Showing: 101 - 120 of 940 results