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We narrowed to 145 results for: transfection

Showing: 21 - 40 of 145 results

  1. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...used by respondents (transfection method, amount of each component per transfection etc.). Take Pooran'...an ssDNA guide. Therefore, researchers must co-transfect cells with 5’-P-ssDNA guides and an NgAgo expression...delivery methods. Researchers can conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA...researchers will have to carry out repeated transfections with the ssDNA guide, although many may find...

  2. 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...

  3. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ... performed 12–18 hours post-transfection to help remove toxic transfection reagents and keep the cells...HEK293T cells, expressing Ad E1A and E1B, can be transfected with your transfer plasmid, carrying your gene...helper plasmids, commonly called “triple plasmid transfection” (Figure 1).    Figure 1: Overview of key steps...cells happy. Harvesting virus Once you have transfected your cells, wait 2–5 days and then collect the supernatant...

  4. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...due to the ease of transduction in difficult-to-transfect cell types ranging from primary to embryonic stem...construct is also reliably higher than traditional transfection and random genomic integration of a plasmid.... is also generally less toxic to cells than transfections, preserving cell viability. Library delivery... with viral vectors instead of a traditional transfection method. This is because viral systems have a...

  5. Antibodies 101: Validation

    Type
    Blog Post
    ...Interest (GOI), and transfect it into your favorite cell line. In parallel, you transfect cells with another...express your protein of interest endogenously and transfect them with a plasmid encoding your protein of interest...expect to see evidence of antibody binding in the transfected cells but not in control cells. But (there is...results show a lovely, strong band from the cells transfected with your protein of interest and show just the... the faintest band from the cells transfected with the other plasmid. Your antibody works! Multiple (Independent...

  6. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ... Options for CRISPR delivery include plasmid transfection, RNP electroporation, and viral transduction... non-replicative. Nanoblades are produced by transfecting the following four components into 293T cells...lower off-target effects when compared to DNA transfection of gRNA and Cas9 expression plasmids. These ...having lower off-target edits compared to DNA transfection.  Homology directed repair (HDR) with Nanoblades...

  7. CRISPR Plasmids - Drosophila

    Type
    Collection
    ... BspQI Transfection Puromycin yes, cut Ji-Long Liu 45946 pU6-BbsI-chiRNA dU6 BbsI Transfection none, need...Bullock and Port 51026 U6-BbsI-crRNA dU6 BbsI Transfection none, need Cas9 plasmid O'Connor-Giles , Harrison...

  8. Genomic Deletions in Mammalian Cell Lines

    Type
    Collection
    ...another transfection method such as cationic liposome-based transfection. Optimize transfection conditions...Perform maxi-prep for each CRISPR/Cas9 construct. Transfecting CRISPRs into Cells of Interest NOTE: This protocol...which is used for MEL cells. However, transient transfection of CRISPR/Cas9 plasmids may be successfully ...types using preferred culture conditions and transfection strategies for each cell type. While MEL cells...Fluorescence Activated Cell Sorting (FACS) of Transfected Cells Prepare cells for FACS by filtering them...

  9. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...Overview: The first step in packaging AAV is transfecting HEK293 cells with AAV packaging plasmids. The...packaged into the virus. Two to five days after transfection, the AAV-containing cells and media are harvested...similar to this one to plan my AAV packaging PEI transfections. It saves me time and helps me figure out if...the PEI calculator for planning AAV packaging transfections Find tips for a first time CRISPR user Resources...

  10. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...we have found that transfecting with PBx mRNA can boost efficiency over transfecting with plasmids in some...traditional method of making stable cell lines is to transfect, select, and isolate clones. This procedure is...integration efficiencies that are accomplished with transfections and are therefore virus-free. This is achieved...fully intact, while, with a standard DNA plasmid transfection, a double-stranded break may occur in your cargo...

  11. TALEN Plasmids and Kits

    Type
    Collection
    ...Gate-assembled TALEN, both in microinjected embryos and transfected cells. TALEN expression is driven by the strong...expression in mammalian systems including hard-to-transfect cell lines. Features include (i) Esp3I (BsmBI)...on pTAL7a and pTAL7b for enrichment of double-transfected mammalian cells. In addition, a T7 promoter was...alternative. Based on transient double-selection of transfected cells, the system has successfully been employed...

  12. Plan Your Experiment

    Type
    Collection
    ...For easy-to-transfect cell types (e.g. HEK293 cells), transfection with standard transfection reagents may...or gRNA in a mammalian cell line that can be transfected at high efficiency Lentiviral transduction Cas9... mammalian cell lines Useful for difficult to transfect cell types, can also be used in vivo A common ...

  13. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ...transient cotransfection of the mARG plamsids into HEK293T cells using a chemical transfection reagent ...proteins. Excess gvpA plasmid added into the transfection mixture in HEK293T cells gave drastic improvement...These plasmids can be used directly in transient transfection in rtTA+ cell lines (such as HEK293-TETOn cells...visible under phase contrast starting day 2 after transfection.     Fig: 2: Expressing mARGAna in MDA-MB...they can also be integrated into the genome by cotransfecting them with a plasmid expressing PiggyBac transposase...saturate expression. When engineering a cell line, cotransfect the mARG cassettes 2:1 (A:N-V) with PiggyBac ...reagent (e.g. PEI-MAX). Simply cotransfect a mixture of 1248 ng of CMV-gvpA-IRES-BFP plasmid and 954 ng CMV-...

  14. 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...

  15. DNA Service - Cloning Grade DNA

    Type
    Collection
    ... different DNA concentrations. Can I directly transfect cloning grade DNA into cells? Cloning grade DNA...designed for use in cloning applications. Direct transfection into cells is not recommended. Can I order more...

  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. CRISPR Plasmids - C. elegans

    Type
    Collection
    ...(see paper) Transfection none, need Cas9 plasmid Calarco 47943 pMB70 cU6 BsaI Transfection none, need ...

  18. Plasmids 101: Protein Expression

    Type
    Blog Post
    ... strategy for protein expression consists of transfecting cells with your DNA template of choice and allowing...SV40, CMV, and RSV) for robust expression post-transfection. Mammalian systems can express proteins both... Both methods produce high protein yields if transfection is successful. Some mammalian systems also ...

  19. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ... how lentiviral particles are produced by co-transfecting three essential ingredients: the lentiviral ...the 3rd-generation system is that it involves transfecting four different plasmids into the producer cells...use lentiviral transfer vectors for transient transfections? A2: Technically, yes. Will it actually work...

  20. Luciferase Plasmid Collection

    Type
    Collection
    ...constitutive promoter and can be used to monitor transfection or viral infection efficiency, to study changes... Renilla luciferase is used to monitor viral transfection Feng Zhang 21471 pLenti PGK V5-LUC Neo (w623... Firefly luciferase is used to moniter viral transfection Sidi Chen 109396 TBG-Vector Firefly TBG AAV-... Firefly luciferase is used to moniter viral transfection Sidi Chen 118412 ssAAV-EF1a-FLuc-WPRE-HgHpA_...
Showing: 21 - 40 of 145 results