Skip to main content
Addgene

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

Information

In Category

We narrowed to 53 results for: PCR-amplified REST

Showing: 41 - 53 of 53 results

  1. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    Published
    Sept. 28, 2017, 2:01 p.m.
    ...inversion creates a template that’s suitable for PCR amplification using pre-designed primers. By using this ...infected the cell type of interest. Repeat rounds of infection and screening by PCR enrich for capsid variants...those that cross the barrier can be identified by PCR later. The PNS presents a different problem: it’s...infect an animal expressing Cre in a cell type of interest, a scientist can isolate DNA from the infected...with increased ability to infect the cell type of interest, which, in the case of Chan et al, was neurons...of a transgenic mouse. 2. Cell-type-specific restricted transgene expression in the CNS and PNS Previously...promoter or enhancer now allows for cell-type-restricted expression in both the CNS (figure 5 in Chan ...

  2. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    Published
    Dec. 2, 2014, 7:23 p.m.
    ...oligoribonucleotide (ORN) interference-PCR (ORNi-PCR) method to suppress PCR amplification of specific DNA sequences...molecules associated with specific genomic regions of interest in a non-biased manner. To achieve this goal, ...

  3. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    Published
    Sept. 10, 2020, 11:45 a.m.
    ...using BsaI, and promoter-gRNA fragments are PCR amplified for cloning into one of three Zeamays codon-...the first gRNA using restriction enzyme BbsI and the second gRNA using restriction enzyme BsaI. If you’... of gRNAs Golden Gate Assembly uses Type IIS restriction enzymes, which cleave outside of their recognition...These expression vectors each contain Type IIS restriction sites flanking the promoter-gRNA construct, but...oligonucleotides. These plasmids contain Type IIS restriction sites that flank the promoter-gRNA constructs...

  4. Plasmids 101: Biotinylation

    Type
    Blog Post
    Published
    Nov. 15, 2018, 1:50 p.m.
    ...Reaction (PCR) by using a biotinylated primer. Biotin can also be enzymatically incorporated during PCR using...complexes bound to your protein of interest leading to amplification of the reporter signal and easier ...macromolecule of interest To utilize the natural binding of biotin and avidin, macromolecules of interest first ...secondary antibody that targets your protein of interest or primary antibody respectively. Avidin bound...sites found on avidin are also ideal for signal amplification during these experiments. During detection, ...methods to chemically biotinylate macromolecules of interest both in vitro and in vivo. Biotinylation reagents...AviTag. Using AviTag, one can label a protein of interest with biotin by simply creating a fusion in which...

  5. 6 Tips for Analyzing and Troubleshooting Sanger Sequencing Results

    Type
    Blog Post
    Published
    June 26, 2014, 2:23 p.m.
    ... requires you to perform your own Big Dye PCR amplification reaction (as opposed to using the all inclusive... from your sequencing runs.   When you run a restriction digest on a gel you always include proper controls...primer at least 50bp upstream of the sequence of interest. 5. Use a silica spin column for purification ...

  6. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...SLIC cloning. Amplify the pCoofy vector and the sequence of gene of interest in separate PCR amplication...during the PCR amplification step, so that only vectors with the desired sequence of interest are retained...method for protein inactivation Your protein of interest does x, but it also participates in y. Depletion...accomplished by knocking down your endogenous protein of interest and co-expressing its replacement recombinant ...into Golden Gate entry vectors via traditional restriction enzyme cloning, TA cloning, or annealing of oligonucleotides...would like to extend this work into their RTK of interest. Grusch, M. et al., EMBO J 2014 Aug 1;33(15)... genetic tag which can be fused to proteins of interest for EM imaging, or targeted to subcellular regions...

  7. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    Published
    June 4, 2020, 1:22 p.m.
    ...degrade the double helix, fragmenting plasmid DNA, PCR product, or chromosome. One of the main causes of...most common E. coli strains used for plasmid amplification are endA negative, meaning the gene has been... tools used in modern molecular biology like restriction enzymes and polymerases. In the situation where...

  8. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    Published
    April 2, 2020, 6:17 p.m.
    ... sequence used as an initiation point for PCR amplification or sequencing. Primers can be exploited for... Short segment of DNA which contains several restriction sites allowing for the easy insertion of DNA....“vectors” or “constructs.” To insert a gene of interest into a vector, scientists may utilize one of a... a variety of cloning methods (restriction enzyme, ligation independent, Gateway, Gibson, etc). The cloning...

  9. A Look at Addgene's QC Process

    Type
    Blog Post
    Published
    May 6, 2025, 1:15 p.m.
    ...places where DNA is being created, such as PCR amplification or bacterial growth. Created with BioRender.com... utilize an array of tools that rely on DNA amplification and manipulation to produce and edit plasmids... sequencing all incoming plasmids. If you are interested in requesting one of these plasmids, you can ...

  10. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    Published
    June 27, 2023, 1:15 p.m.
    ... the same pathways. Additionally, as GPCR signaling is amplified in successive steps along a pathway, ...controlling GPCR activation have been developed to selectively study individual types of GPCRs with greater...be used for lower frequency stimulation. GPCR signaling GPCR signaling is a complicated process involving...change the membrane potential. OptoXRs are modified GPCRs. When exposed to light, they activate downstream... coupled receptors  G-protein coupled receptors (GPCR) regulate cellular function through multiple intra-cellular...have been developed for utilizing light to control GPCR activation and downstream signaling cascades (reviewed...Levitz, 2020), including light-sensitive chimeric GPCRS (OptoXRs) (Tichy et al., 2019), photoswitchable ...

  11. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    Published
    March 15, 2016, 2:30 p.m.
    ...you to quantify the amount of PCR product that comes from amplification of the original plasmid which ... titer are direct p24 measurements via ELISA and qPCR for viral RNA. Measuring p24 levels in a viral prep... based off of p24 quantification tend to be overestimates since they can include free p24 and defective...first converted to cDNA and then quantified using qPCR primers targeting specific viral components such...assay, titering via measurement of viral RNA can overestimate the amount of infectious virus in a prep as ...assays depending on the vector backbone. When using qPCR based methods for viral titration consider using...physical titer, there is a risk that they may underestimate viral titer as these assays cannot distinguish...

  12. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    Published
    May 10, 2016, 2:30 p.m.
    ... along with Cas9 in the T-cell line 2D10. PCR amplification and Sanger sequencing of pooled samples showed...viral protein production, so researchers are also interested in precisely excising the HIV-1 genome from infected...

  13. Hot Plasmids: Winter 2025

    Type
    Blog Post
    Published
    March 11, 2025, 1:15 p.m.
    ...activation kinetics and high responsivity near the resting membrane potential, ASAP5 is a great new option... an enhanced pU6cm promoter, viral replicon amplification of cargo, and heat treatment.  Figure...CLAP) with a flexible linker where a protein of interest can be inserted. B) Protein expression and labeling...studies of kinesins but for many other proteins of interest too! Find CLIP, CLAP, and SNAP plasmids here! ...biosensors kit By Mike Lacy G protein-coupled receptors (GPCRs) are critical components of many physiological pathways...targets. To enable large-scale profiling of many GPCRs, Mikel Garcia-Marcos’ lab recently developed a collection... detection of activation triggered by endogenous GPCR. Example applications include large-scale pharmacological...
Showing: 41 - 53 of 53 results