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Showing: 1 - 15 of 15 results

  1. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...' cap structure, and proceeds through U5 and the rest of the provirus. Some newer plasmids use a hybrid...plasmid — contains transgene, sgRNA, or shRNA of interest flanked by LTRs; ~8 kb packaging capacity Packaging...Cloning Cloning your transgene, gRNA, or shRNA of interest into the transfer plasmid can be done with most...most standard cloning methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer...transfer plasmids may have limited restriction sites or may only be compatible with certain cloning methods ...virus can then be used to transduce the cells of interest, either with or without determining the viral ...components. The transfer plasmid encodes the gene of interest and contains the sequences that will be incorporated...

  2. Adenovirus Guide

    Type
    Guide
    ...adenoviral plasmid. Adenoviral plasmid — containing the rest of the adenoviral genome necessary for viral production...transgene of interest is cloned into the shuttle plasmid, verified, and linearized with the restriction enzyme...regions between the two ITRs with the transgenes of interest. The size of the genetic cargo that can be included...transfer plasmid) — containing the transgene of interest, the ITRs, and homology regions for recombination...Cloning Cloning your transgene, gRNA, or shRNA of interest into the transfer plasmid can be done with most...most standard cloning methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer...transfer plasmids may have limited restriction sites or may only be compatible with certain cloning methods ...

  3. Lentiviral Vector Guide

    Type
    Guide
    ...of R, is capped, and proceeds through U5 and the rest of the provirus. Third-generation plasmids use a...plasmid — contains transgene, sgRNA, or shRNA of interest flanked by LTRs; 8–10 kb packaging capacity Packaging...Cloning Cloning your transgene, gRNA, or shRNA of interest into the transfer plasmid can be done with most...most standard cloning methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer...transfer plasmids may have limited restriction sites or may only be compatible with certain cloning methods ...concentrated viral particles to transduce the cells of interest, either with or without determining the viral ...plasmids. The transfer plasmid encodes the gene of interest and contains the sequences that will be incorporated...

  4. Cloning

    Type
    Guide
    ... In restriction cloning, scientists utilize specific restriction enzymes to cut dsDNA of interest into...used to create recombinant DNA. Restriction Enzyme Cloning Restriction enzyme (endonuclease) based molecular... find a protocol for restriction cloning and an in-depth breakdown of restriction digests on our website...website. Restriction cloning of your gene of interest (YGOI) into a recipient plasmid. (Image from Plasmid...Empty Backbone Guide Genome Engineering Guide Restriction Enzyme Gateway TOPO Gibson Type IIS Ligation ...scientists can amplify and manipulate genes of interest and then insert them into plasmids for replication...reasons, remains one of the most popular today. Restriction enzymes, which are naturally produced by certain...

  5. Antibody Guide

    Type
    Guide
    ...protein of interest. Signal strength is determined by how many antibodies bind the protein of interest, while... antibody of interest to a second, independent antibody for the same protein of interest, with no overlapping...quantify, identify, separate, and locate proteins of interest. The protein that an antibody recognizes is called... of the animal’s immune system. The antigen of interest is introduced to the animal, which then mounts...primary antibody which is bound to your protein of interest. Secondary antibodies can be isotype-specific ...recognize an available epitope on your protein of interest. Polyclonal antibodies are typically collected...Western blots should be run with your protein of interest and at least one ubiquitously expressed protein...

  6. Molecular Biology Reference

    Type
    Guide
    ...contains several restriction enzyme sites, enabling easy insertion of DNA by restriction enzymes digestion...used for cloning by restriction enzyme digestion. For more information about restriction enzymes check out...human genome. Digesting the human genome with restriction enzymes would yield about two million DNA fragments...plasmid. By the 1970s the combined discoveries of restriction enzymes, DNA ligase, and gel electrophoresis ...molecule, which can be used to study DNA fragments of interest, such as genes. Creation of recombinant DNA Molecular...antibiotic-resistance gene, and at least one unique restriction enzyme recognition site. These elements allow... not carrying the plasmid. Additionally, the restriction enzyme site(s) allow for the cloning of a fragment...

  7. Plan Your Experiment

    Type
    Guide
    ...nucleotide edits that introduce a novel restriction site. The region of interest is PCR amplified, digested with...region of interest can be PCR amplified using primers that (A) flank the region of interest (deletions...necessary to model your specific disease or process of interest. Do you want to: Generate complete and permanent...the promoter driving expression of your gene of interest. For genetic knockouts, gRNAs commonly target ... containing the gRNA scaffold using standard restriction-ligation cloning. However, the exact cloning ... within a mixed cell population. The region of interest is PCR amplified, PCR products are denatured-renatured...agarose gel to identify DNA fragments. PCR and restriction digest (for HDR repaired DSBs): For small nucleotide...

  8. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...cassette that typically contains the transgene of interest. The size of the genetic cargo that can be included... as cis plasmid) — containing the transgene of interest between the two ITRs. Packaging plasmid (also ...Cloning Cloning your transgene, gRNA, or shRNA of interest into the transfer plasmid can be done with most...most standard cloning methods, including restriction enzyme, Gibson Assembly, or Gateway. Some transfer...transfer plasmids may have limited restriction sites or may only be compatible with certain cloning methods ...fine-tune tropism towards specific cell types of interest has been to create and test capsid libraries with...biological structures. Specifically, researchers were interested in creating systemic capsids that could target...

  9. Chemogenetics Guide

    Type
    Guide
    ... Human M3 muscarinic Arrestin-2/-3 CNO* Increase Arrestin translocation Arrestin signalling Nakajima &...used. There is also a DREADD that couples to β-arrestin to activate noncanonical GPCR signaling independent...used to target AAV vectors to specific cells of interest. In addition, different AAV serotypes have varying...Design and functional characterization of a novel, arrestin-biased designer G protein-coupled receptor. Mol... Stem Cell-Derived Neurons Repair Circuits and Restore Neural Function. Cell Stem Cell. 28:112-126.e6....

  10. CRISPR Guide

    Type
    Guide
    ...conditions: The sequence is unique compared to the rest of the genome The target is present immediately ...Publications CRISPR Overview Bacteria have an interesting adaptive immune response that detects and eliminates... Cas enzyme and a gRNA specific to the gene of interest. The genomic target of the gRNA can be any ~20...activity with endogenous cell-type specific miRNAs to restrict CRISPR editing only to the target tissue. Browse... molecules associated with a genomic region of interest in vivo. Figure 12: Overview of purifying genomic...

  11. Guide to Using Pooled Libraries

    Type
    Guide
    ... the majority of the genome, while others are restricted to certain gene sets. In cDNA libraries, each... DNA can be delivered directly to the cells of interest. With others, notably pooled lentiviral plasmid...subsequently used to deliver the plasmids to the cells of interest. In either case, next-generation sequencing of...

  12. Educational Resources

    Type
    Guide
    ...Learn about plasmids, sequencing, vectors, and restriction enzymes to assist with your molecular biology...Plasmid Cloning Creating Bacterial Glycerol Stocks Restriction Digests Guides Browse our in-depth instructional...

  13. Science Guides

    Type
    Guide
    ...to capture, quantify, and identify proteins of interest. Read More Chemogenetics Chemogenetics is a technique... the majority of the genome, while others are restricted to certain gene sets. Read More Viral Vectors...

  14. Promoters

    Type
    Guide
    ...controls when and where in the organism your gene of interest is expressed. Summary Promoters are about 100-...

  15. Optogenetics Guide

    Type
    Guide
    ...utilized to control the localization of a protein of interest. A genetically-encoded nuclear Cry2 fusion can...
Showing: 1 - 15 of 15 results