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We narrowed to 457 results for: nsf

Showing: 161 - 180 of 457 results

  1. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ...most cases, interested in transferring genes to mammalian cells (i.e. transferring DNA), yet retroviruses...generalize the idea of “transduction” to mean any transfer of material from the virus to the host. In fact...pseudotransduction” — to describe the direct protein transfer of RT, which would actually occur in step 2 during...

  2. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...brain? Cells of a specific brain area can be transfected by in utero electroporation. To achieve sparse... could reduce the plasmids amounts to reduce transfection efficiency. However, if the cellular structure... it reduces the probability that neurons are transfected with both vectors. Flex vectors are the solution... FLEx vectors guarantees that most cells are transfected with all labeling plasmids and the low concentration...

  3. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...others opt for a single harvest after transfecting with the transfer and packaging plasmids. A benefit of...the passage number of a culture the lower the transfection and transduction efficiencies. Cultures should...multiplicity of infection (MOI) for predicting gene transfer events.” Genetic Vaccines and Therapy 2.1 (2004...

  4. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ... More information on Agrobacterium-mediated transformation can be found in this blog post. The pDGE Dicot...Simplified schematic of agrobacterium-mediated transformation in plant cells. Agrobacterium tumefaciens can...plant genome. B) The Ti plasmid and helper are transformed into agrobacterium tumefaciens and exposed to...Gelvin SB (2003) Agrobacterium-Mediated Plant Transformation: the Biology behind the “Gene-Jockeying” Tool...

  5. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...methylation-dependent endonuclease (i.e. DpnI), and bacteria are transformed with the nuclease-resistant nicked plasmid (the...the adenosine of the GATC recognition site. Transformation: After the PCR reaction, no ligation is required...required since the E. coli you transform your PCR products into will efficiently patch up the DNA. The ...parental plasmid provides a mean for selecting for transformants which have taken up the mutagenized plasmid....

  6. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...process known as bioluminescent resonance energy transfer (BRET). The fluor then emits light at a tissue-penetrating...have been made using fluorescent resonance energy transfer (FRET) probes. These probes generally consist ...increasing their FRET signal. FRET is a non-radiative transfer of energy from one fluorescent protein to a compatible...activation of other fluorescent probes but can still transfer energy to the fluorescent protein acceptor. NLuc...

  8. Streaking for Single Colonies: The Streak Plate Challenge

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    Blog Post
    ...students transform them into different strains of E. coli,” says Westenberg. During the transformation experiment...genetic backgrounds of the host strains affect transformation efficiency.  Aside from the Streak Plate Challenge...

  9. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...interference (RNAi). The most common method is transfection of short interfering RNA (siRNA). Knockdown .... This approach often involves the transient transfection of expression plasmids in a given cell line....antibody should be able to detect even in cells transfected with low efficacy. But since there can be large...differences between endogenous expression and transfected overexpression, this approach cannot confirm ...

  11. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    ...enables selection of cells with the plasmid post transfection. The 2A "self cleaving" peptide causes the ribosome...backbone. This particular backbone can be used to transfect a variety of mammalian cells and can be modified...can be used to test for SpCas9 expression post transfection by western blot. Overall, this is a versatile...

  12. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    ...system which has been designed to perform gene transfer in vertebrates. The system is comprised of 2 components...the lab showed that efficiencies of stable gene transfer using SB100x were comparable to stable transduction... Beauty transposase enables robust stable gene transfer in vertebrates. Nat Genet. 2009 Jun;41(6):753-...

  13. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ...mammalian selection cassette to screen for positive transfected cells. The addition of an in-frame fluorescent...you can get away with generating transiently transfected cells, you will not need a mammalian selection...create a new strain with a disrupted gene by transforming wild-type yeast with an integrating vector expressing...

  14. Plasmids 101: Origin of Replication

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    Blog Post
    ...plasmids from the same group should not be co-transformed, so if you need two plasmids for an experiment... same incompatibility group should not be co-transformed. +Actual copy number varies. See below for ...stocks, agar stabs, or liquid cultures. Freshly transformed cells also give a higher plasmid yield that colonies...

  15. "Build Your Plasmid": The Game - Play to Teach

    Type
    Blog Post
    ...Resistance gene and Selection Markers) and one “Transfer Plasmid” game board. Once they agreed on each ...had 35 minutes to develop a strategy and build a transfer plasmid that could be used to genetically manipulate... this way, the students also had to practice transferable skills like team building, collaborative critical...

  16. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    ..., you’re ready to edit. These cells are then transfected with several different constructs for the reporter... gRNAs by simply changing the gRNA used when transfecting your cells.  A plasmid that contains Cas9. The...plasmid.  With this set up, all successfully transfected cells will express RFP while cells containing...

  17. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    ...pSpCas9(BB)). The 2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP (PX458). lentiCRISPR...lentiCRISPR v2 - This 3rd generation lentiviral transfer vector also from the Zhang lab is used to deliver... David Root lab is a 3rd generation lentiviral transfer plasmid for cloning and expressing new shRNA sequences...

  18. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    ...DNA molecule as the gene that they package. In a transfer plasmid, the transgene is flanked by two ITRs,...and rAAV vectors, modifications can affect gene transfer efficiency and transgene expression (Zhou et al...of ITR and capsid serotypes vary in their gene transfer efficiency. Our practical knowledge of ITRs is...

  19. Viral Vectors 101: Viral Vector Elements

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    Blog Post
    ...the delivered cargo (often a gene) is called the transfer plasmid while the other plasmids with packaging... or something similar, and is contained on the transfer plasmid. If expression is the goal, a promoter...delivered and expressed in the host cell from the transfer plasmid. Viral packaging and entry The viral packaging...

  20. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    ...DNA. The other component of these systems, methyltransferases, methylate DNA at particular sequences to...restriction modification systems containing methyltransferases that add methyl groups to specific DNA sequences...plasmids with no methylation (there are no methyltransferases in the PCR reaction). The template plasmid...
Showing: 161 - 180 of 457 results