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We narrowed to 48 results for: gag pol vector

Showing: 21 - 40 of 48 results
  1. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...replication. They contain two genes: gag and pol. The pol polyprotein encodes the reverse transcriptase ...in viral vectors, but easily fit in a TE. Transposons are also less likely than viral vectors to induce...transposon’s LTRs, similar to when packaging viral vectors.   Let’s take a closer look at three different ... in human cells at rates similar to retroviral vectors. The hyperactive version of the SB transposase,...efficiency in human cells similar to retroviral vectors. The hyperactive PB transposase (hyPB) is ~10-fold...transfer Transposons are an alternative to viral vector delivery of transgenes, like for iPSC reprogramming... piggyBac, which is a big advantage over viral vectors (~5 kB payload for AAV and ~8 kB payload for lentivirus...
  2. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...replication. They contain two genes: gag and pol. The pol polyprotein encodes the reverse transcriptase ...in viral vectors, but easily fit in a TE. Transposons are also less likely than viral vectors to induce...transposon’s LTRs, similar to when packaging viral vectors.   Let’s take a closer look at three different ... in human cells at rates similar to retroviral vectors. The hyperactive version of the SB transposase,...efficiency in human cells similar to retroviral vectors. The hyperactive PB transposase (hyPB) is ~10-fold...transfer Transposons are an alternative to viral vector delivery of transgenes, like for iPSC reprogramming... piggyBac, which is a big advantage over viral vectors (~5 kB payload for AAV and ~8 kB payload for lentivirus...
  3. Hot Plasmids: Spring 2025

    Type
    Blog Post
    Published
    June 3, 2025, 1:15 p.m.
    ...fusions for membrane display, they recommend the NGR vectors for most experiments. These new tools promise to... toolkit is the empty plasmid backbone: the DNA vector that contains everything needed to replicate inside...composed of a protein core, 2–4 glycosaminoglycans (GAG), and are (typically) tethered to the cell surface...successful experiments or therapies. Using AAV vectors with enhancers that drive cell-type-specific expression...colleagues characterized a series of new enhancer-AAV vectors targeting specific classes of cortical and striatal...Modular by design: Key elements (promoter, tag, polyA site) are flanked by unique restriction sites so...general heat-shock proteins using IP with CycT1 or PolII-CTD antibodies and unlabeled MS in DLD-1 cells (...
  4. Hot Plasmids: Spring 2025

    Type
    Blog Post
    Published
    June 3, 2025, 1:15 p.m.
    ...fusions for membrane display, they recommend the NGR vectors for most experiments. These new tools promise to... toolkit is the empty plasmid backbone: the DNA vector that contains everything needed to replicate inside...composed of a protein core, 2–4 glycosaminoglycans (GAG), and are (typically) tethered to the cell surface...successful experiments or therapies. Using AAV vectors with enhancers that drive cell-type-specific expression...colleagues characterized a series of new enhancer-AAV vectors targeting specific classes of cortical and striatal...Modular by design: Key elements (promoter, tag, polyA site) are flanked by unique restriction sites so...general heat-shock proteins using IP with CycT1 or PolII-CTD antibodies and unlabeled MS in DLD-1 cells (...
  5. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT vector ensures...science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector backbone with ...custom-cloned vectors in a Multisite Gateway reaction with a desired destination vector expressing a separate...a final lentiviral vector containing up to 5 different elements. Lentiviral vectors constructed with the...MXS-chaining kit contains 5 empty chaining vectors plus a destination vector and a collection of building blocks...complex polycistronic lentiviruses. The MuLE toolbox contains a series of building-block vectors that are.... Additionally, by using an adapted destination vector with a heterologous promoter outside of the Multisite...
  6. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...expression vector which gives close to 100% cloning efficiency and gene expression. The pUCXKT vector ensures...science.aaa3380 MuLE vectors for customized lentivirus production Finding a lentiviral vector backbone with ...custom-cloned vectors in a Multisite Gateway reaction with a desired destination vector expressing a separate...a final lentiviral vector containing up to 5 different elements. Lentiviral vectors constructed with the...MXS-chaining kit contains 5 empty chaining vectors plus a destination vector and a collection of building blocks...complex polycistronic lentiviruses. The MuLE toolbox contains a series of building-block vectors that are.... Additionally, by using an adapted destination vector with a heterologous promoter outside of the Multisite...
  7. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ...of pExpression vectors) plasmids. The type IIS restriction sites used in these vectors are of low abundance...to produce the constructs you need. Expression vectors within this kit contain a variety of promoters ...insects, and even Xenopus oocytes. The expression vectors additionally contain various combinations of protein...Series Confronted with the problem of creating vectors for predictable protein expression in bacteria,...this so-called bicistronic design (BCD). The 49 vectors in this kit can be used to drive production of ...SITS to create the pCell-Free vectors, Gateway-compatible backbones that enable cell-free expression of ...kit represents a set of 88 clones in a pCellFree vector that enables protein expression in any in vitro...
  8. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ...of pExpression vectors) plasmids. The type IIS restriction sites used in these vectors are of low abundance...to produce the constructs you need. Expression vectors within this kit contain a variety of promoters ...insects, and even Xenopus oocytes. The expression vectors additionally contain various combinations of protein...Series Confronted with the problem of creating vectors for predictable protein expression in bacteria,...this so-called bicistronic design (BCD). The 49 vectors in this kit can be used to drive production of ...SITS to create the pCell-Free vectors, Gateway-compatible backbones that enable cell-free expression of ...kit represents a set of 88 clones in a pCellFree vector that enables protein expression in any in vitro...
  9. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    Published
    June 27, 2023, 1:15 p.m.
    ...concentration gradient and hyperpolarize the cell (Mattis et al., 2012). Bicistronic vectors combine excitation...candidates for your experiments.  Wavelengths Most vectors have an optimal wavelength to use but can be activated...enter the cell with the concentration gradient to depolarize it (Nagel et al., 2003). Inhibitory opsins are...the receptor, when exposed to light, the ligand engages the binding pocket and activates the receptor. ...considering for your experiment, note that you cannot extrapolate results from one cell type to another, as they... A. K., Hegemann, P., & Wiegert, J. S. (2021). BiPOLES is an optogenetic tool developed for bidirectional...
  10. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    Published
    June 27, 2023, 1:15 p.m.
    ...concentration gradient and hyperpolarize the cell (Mattis et al., 2012). Bicistronic vectors combine excitation...candidates for your experiments.  Wavelengths Most vectors have an optimal wavelength to use but can be activated...enter the cell with the concentration gradient to depolarize it (Nagel et al., 2003). Inhibitory opsins are...the receptor, when exposed to light, the ligand engages the binding pocket and activates the receptor. ...considering for your experiment, note that you cannot extrapolate results from one cell type to another, as they... A. K., Hegemann, P., & Wiegert, J. S. (2021). BiPOLES is an optogenetic tool developed for bidirectional...
  11. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ...oligo when cloning my target spacers into the PX330 vector backbone? A2: There is no need to add the NGG PAM...recombination (HR) Q7: Can you summarize HR template vector design guidelines? A7: A few notes below are considerations...recently is the double nickase system, with PX335 vector, you need to find two guides for each cleavage ... co-deliver into the cells with the Cas9/sgRNAs vector (like PX330). For large changes (>100 bp insertions...typical size (~5 kb) donor plasmid with Cas9/sgRNAs vector, for one well in 24-well plate. A good example/...typical size (~5 kb) donor plasmid with Cas9/sgRNAs vector, for a one well in a 24-well plate. A good reference... and surveyor. Q15: I used DNA polymerase Takara Ex Taq™ DNA Polymerase for my genomic PCR, but couldn't...
  12. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ...oligo when cloning my target spacers into the PX330 vector backbone? A2: There is no need to add the NGG PAM...recombination (HR) Q7: Can you summarize HR template vector design guidelines? A7: A few notes below are considerations...recently is the double nickase system, with PX335 vector, you need to find two guides for each cleavage ... co-deliver into the cells with the Cas9/sgRNAs vector (like PX330). For large changes (>100 bp insertions...typical size (~5 kb) donor plasmid with Cas9/sgRNAs vector, for one well in 24-well plate. A good example/...typical size (~5 kb) donor plasmid with Cas9/sgRNAs vector, for a one well in a 24-well plate. A good reference... and surveyor. Q15: I used DNA polymerase Takara Ex Taq™ DNA Polymerase for my genomic PCR, but couldn't...
  13. Plasmid Cloning by PCR

    Type
    Blog Post
    Published
    March 29, 2016, 2:30 p.m.
    ...will describe how to copy a cDNA from one vector into a new vector that is better suited for analyzing the...use a high fidelity polymerase to minimize mutations. The fidelity of the polymerase becomes more important... million bp depending on the polymerase used. No matter which polymerase you use, it is important that.... This gives us a sequence of 5'-TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3' (30bp with 18bp of homology to...chose for our reverse primer (5’-TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3’) into this calculator we get a...final Reverse Primer sequence of 5’-TGCTTAGCGGCCGCTCAGTACTTCGAGATATGCCA-3’. Preparing the PCR product for...
  14. Plasmid Cloning by PCR

    Type
    Blog Post
    Published
    March 29, 2016, 2:30 p.m.
    ...will describe how to copy a cDNA from one vector into a new vector that is better suited for analyzing the...use a high fidelity polymerase to minimize mutations. The fidelity of the polymerase becomes more important... million bp depending on the polymerase used. No matter which polymerase you use, it is important that.... This gives us a sequence of 5'-TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3' (30bp with 18bp of homology to...chose for our reverse primer (5’-TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3’) into this calculator we get a...final Reverse Primer sequence of 5’-TGCTTAGCGGCCGCTCAGTACTTCGAGATATGCCA-3’. Preparing the PCR product for...
  15. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ...reprogramming discrepancies. The polycistronic piggyBac transposon vectors tested in this work are available...TagRFPt, EGFP, mVenus, mCerulean3, mKOFP2) and empty vectors that have standard cloning sites or gateway compatible...neuronal tools to the expanding gateway compatible vectors currently shared amongst the zebrafish community...for the research community. You can find the AAV vectors for infecting GABA-ergic interneurons here. Dimidschstein...Very low off-target activity; protein purification vector  87439  pCMV-HF-BE3  HF-Cas9 (NGG)  Very low ...low off-target activity; mammalian expression vector Rees H, et al. Nature Communications. 2017. PubMed...editing. The bacterial and mammalian expression vectors for these CRISPR inhibitors are listed in the table...
  16. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ...reprogramming discrepancies. The polycistronic piggyBac transposon vectors tested in this work are available...TagRFPt, EGFP, mVenus, mCerulean3, mKOFP2) and empty vectors that have standard cloning sites or gateway compatible...neuronal tools to the expanding gateway compatible vectors currently shared amongst the zebrafish community...for the research community. You can find the AAV vectors for infecting GABA-ergic interneurons here. Dimidschstein...Very low off-target activity; protein purification vector  87439  pCMV-HF-BE3  HF-Cas9 (NGG)  Very low ...low off-target activity; mammalian expression vector Rees H, et al. Nature Communications. 2017. PubMed...editing. The bacterial and mammalian expression vectors for these CRISPR inhibitors are listed in the table...
  17. The time and cost required to make a plasmid

    Type
    Blog Post
    Published
    March 23, 2023, 1:15 p.m.
    ...restriction enzymes or vector purchases (Table 1). Restriction enzymes and vector purchases can raise the...for the simple insertion of a PCR product into a vector. The time it takes to make a plasmid could increase...plasmid? Figure 2: Survey results from a Twitter poll run in 2023, n=194.   After the great Plasmid...much harder to calculate, in both our 2018 and 2023 poll, most people said it took them more than 22 hours...these rough estimates, it’s clear that community engagement with and use of repositories like Addgene provides...
  18. The time and cost required to make a plasmid

    Type
    Blog Post
    Published
    March 23, 2023, 1:15 p.m.
    ...restriction enzymes or vector purchases (Table 1). Restriction enzymes and vector purchases can raise the...for the simple insertion of a PCR product into a vector. The time it takes to make a plasmid could increase...plasmid? Figure 2: Survey results from a Twitter poll run in 2023, n=194.   After the great Plasmid...much harder to calculate, in both our 2018 and 2023 poll, most people said it took them more than 22 hours...these rough estimates, it’s clear that community engagement with and use of repositories like Addgene provides...
  19. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ...domains of the selected scFvs are inserted into a vector encoding the antibody constant domain scaffold ...small size also allows for their delivery by viral vectors like AAV, which could be useful for delivering ...doi.org/10.1038/nbt.1791  Deal CE, Balazs AB (2015) Vectored antibody gene delivery for the prevention or treatment...recombinant antibody. They are ~25 kDa single polypeptides that contain the variable light chain (VL) and...When these molecules, called Bispecific T-cell engagers (BiTE®s), bind CD3 on T cells and a tumor-specific...
  20. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ...domains of the selected scFvs are inserted into a vector encoding the antibody constant domain scaffold ...small size also allows for their delivery by viral vectors like AAV, which could be useful for delivering ...doi.org/10.1038/nbt.1791  Deal CE, Balazs AB (2015) Vectored antibody gene delivery for the prevention or treatment...recombinant antibody. They are ~25 kDa single polypeptides that contain the variable light chain (VL) and...When these molecules, called Bispecific T-cell engagers (BiTE®s), bind CD3 on T cells and a tumor-specific...
Showing: 21 - 40 of 48 results