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We narrowed to 134 results for: plasmid transfection

Showing: 41 - 60 of 134 results

  1. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...consisted of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated...and host genome integration. After co-transfection of these plasmids, 293T cells released transgene-containing... packaging and envelope plasmids are two of Addgene’s most requested plasmids! Lentiviral particles with...Many CRISPR plasmids are designed for lentiviral use, including Addgene’s top requested plasmid in 2015: ...itself due to a few missing proteins. The envelope plasmid contained a viral envelope to dictate the tropism...the vector could infect. Finally, the transfer plasmid encoded the desired transgene flanked by HIV-1 ...recombination between the transfer and packaging plasmids does exist. To further reduce this possibility...

  2. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...three plasmids — a transfer plasmid containing your gene of interest plus two packaging plasmids (three...baculovirus plasmid. Production of baculovirus requires only two plasmids — your transfer plasmid and the ...three packaging plasmids for 3rd generation lentivirus production). The packaging plasmids are widely available...to worry about cloning your transfer plasmid. The transfer plasmids have different packaging capacities...in mammalian cells is relatively quick. After transfection, HEK-293T cells start producing virus within...systems, it is standard to do only one passage (transfection). This differs from typical insect cell preparations...called a bacmid. Like AAV and lentiviral packaging plasmids, bacmids are readily available, so the amount ...

  3. Viral Production at Addgene

    Type
    Blog Post
    ...Pennsylvania Vector Core. Transfections are performed using the transfer plasmid, a plasmid encoding rep and serotype-specific...with primers targeting the transfer plasmid used in the transfection. PCR products are analyzed by gel .... coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ... serotype-specific cap genes, and a plasmid encoding adenoviral helper sequences. AAV preparations are...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...offline and discarded. Confirmation of the transfer plasmid Addgene uses a rigorous barcode matching system...microbial contamination. Purity When possible, all plasmids used for viral production are propagated in the...

  4. CRISPR Plasmids - Drosophila

    Type
    Collection
    ... Cas9 plasmid Bullock and Port 51026 U6-BbsI-crRNA dU6 BbsI Transfection none, need Cas9 plasmid O'Connor-Giles...CRISPR Plasmids: Drosophila Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime...gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter...with a wide variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance... none, need Cas9 plasmid Bullock and Port 49330 pAc-sgRNA-Cas9 dU6 BspQI Transfection Puromycin yes, cut...Liu 45946 pU6-BbsI-chiRNA dU6 BbsI Transfection none, need Cas9 plasmid O'Connor-Giles , Harrison , Wildonger...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...

  5. Genomic Deletions in Mammalian Cell Lines

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    Collection
    ...usage of the pSpCas9(BB) plasmid (pX330) (Addgene plasmid ID 42230). This plasmid allows for the simultaneous...another transfection method such as cationic liposome-based transfection. Optimize transfection conditions...utilized, such as pX458 (Addgene plasmid ID 48138) or pX459 (Addgene plasmid ID 48139), which include GFP ...for MEL cells. However, transient transfection of CRISPR/Cas9 plasmids may be successfully adapted to ... with a PCR cloning kit) into a plasmid vector. Transform the plasmid into DH5α E. coli cells and plate...Mammalian Cell Lines Other Addgene Resources CRISPR Plasmids CRISPR Guide Addgene Blog Posts Addgene is proud...multiple sgRNAs may be expressed from a single plasmid. Add “CACC” before the 20-mer guide sequence and...

  6. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...approach often involves the transient transfection of expression plasmids in a given cell line. For flow cytometry...interference (RNAi). The most common method is transfection of short interfering RNA (siRNA). Knockdown ... (tagged or untagged) protein When expression plasmids are available, or the necessary expertise to clone...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 ...

  7. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...can be produced through a single or dual-plasmid transfection, depending on whether the heavy and light...contained to one plasmid or if they’ve been separated onto two. You’ll first transfect your cells with ... great collection of ready-to-use rAbs and rAb plasmids!). Since recombinant antibodies conveniently allow...with your plasmid(s), and then wait 7-14 days (in mammalian cells) to collect the cell culture supernatant...on the Addgene blog Introduction to Antibodies Plasmid-based Recombinant Antibodies Polyclonal Antibodies...

  8. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ... target plasmid and pX330-sgRNA plasmid (Mashiko et al. 2013). The pCAG-EGxxFP target plasmid contains... (c) The pCAG-EGxxFP target plasmid was co-transfected with pX330 plasmids containing sgRNA sequences ... simple matter of transfecting HEK293T cells with your modified pCAG-EGXXFP plasmid along with the individual...individual gRNAs in their PX330 plasmids (or similar plasmids for gRNA expression), one gRNA at a time.... system floating around, and many pre-designed plasmids containing these variants ready for purchase. ...multi-cloning site (MCS) between EGFP fragments. The pX330 plasmid contains humanized Cas9 expression cassette and... you can insert this region into a pCAG-EGXXFP plasmid using standard cloning techniques (The XX indicates...

  9. TALEN Plasmids and Kits

    Type
    Collection
    ...List of plasmid kits and accessory plasmids available from Addgene for generating your own specific TALE...Yamamoto Additional Plasmids for Use with Addgene TALEN Kits For Use With: ID Plasmid Depositing PI Description...destination plasmids were optimized for TALEN expression in mammalian systems including hard-to-transfect cell...TALE Toolbox 49622 - 49647 24 plasmids Thoru Pederson TALEColor plasmids: destination vectors for use ...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail... TALE plasmids. Genome... Genome Engineering TALENs TALEN Plasmids and Kits You may also like... Addgene's Genome...

  10. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    ...maintained DNA plasmids that express reprogramming factors To date, the mRNA transfection method has not...and to model diseases in vitro. Find stem cell plasmids at Addgene Many sources of adult somatic cells...generating hiPSCs for clinical purposes: mRNA transfection- this allows transient expression of the reprogramming...group is an optimized mixture of three episomal plasmids (Table 1) encoding the reprogramming factors. ...enhance the reprogramming efficiency, an extra plasmid, pCXWB-EBNA-1 is used in the reprogramming cocktail...increases protein expression from the other episomal plasmids. When used on PBMCs, the pCXLE episomal vector...available at Addgene. Table 1: List of pCXLE episomal plasmids, their Addgene ID numbers and the reprogramming...

  11. DNA Service - Cloning Grade DNA

    Type
    Collection
    ...Grade DNA (cgDNA) for over 200 plasmids in our collection. The plasmids available in this format include...variety of popular plasmids and backbones with high cloning potential. By making these plasmids available as...individual plasmid page to find current pricing information Standard Quality Control Plasmid verified by...next generation sequencing Plasmids Available as Cloning Grade DNA ID Plasmid Description Industry PI FAQ...grade DNA preps of this plasmid available, but helps to inform us of which plasmids would be most helpful... our most popular plasmids! DNA... DNA Service: Cloning Grade DNA Service Details Plasmids Available FAQ Addgene...

  12. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    ...This is straightforward in easy-to-transfect cell lines where plasmid dosage can be tightly controlled....because it relies on a single BiFC pair. Find BiFC plasmids for your experiment at Addgene! How to Design ...BiFC constructs are expressed transiently by transfection or stably impacts the success of BiFC experiments...Resources on Addgene.org Find fluorescent protein plasmids and resources Visit our Molecular Biology Reference...

  13. Plan Your Experiment

    Type
    Collection
    ...design. Addgene offers many plasmids containing validated gRNAs . These plasmids contain gRNAs that have ...into mammalian cells is to directly transfect expression plasmids encoding your Cas protein and gRNA, ...generate stable cell lines if your plasmid contains a selection marker. Plasmid delivery is versatile thanks...efficiently transfect. However, note that this method is most useful for cell lines that you can transfect at ...cells. These methods are more transient than plasmid transfection or viral vectors, reducing the risk of accumulating...difficult to transfect, transduce, or may not express common promoters. Browse Cas9 plasmids for RNP delivery...Application Cas Enzyme Additional Considerations Browse Plasmids Knockout Permanently disrupt gene function; cells...

  14. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...comparison of an shRNA plasmid sequence and an miRNA plasmid sequence. (A) Plasmid sequence from pLVUHshp53... small RNA and can occur via transfection or viral delivery of a plasmid. RNAi is an umbrella term for....   Expressing shRNAs using plasmids The design of shRNA plasmids requires three important steps: ...between the red boxes. (B) Plasmid sequence from pcDNA3-miR21 (Addgene plasmid #21114). The miRNA sequence...via transient transfections and are handy for quick in vitro experiments. shRNA plasmids typically use...shRNA plasmids are being designed using RNA polymerase II promoters. The design of these plasmids is meant...with Cas13a (C2c2) Plasmids 101: The Promoter Region – Let's Go! Browse All Plasmids 101 Posts Additional...

  15. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ... create fusions with a gene of interest. These plasmids will be most useful for constructing intermolecular...ratio of acceptor to donor fusions varies with transfection efficiency and any unpaired fluorescent proteins...measure (overexpressing a gene or protein after transfection). Future of FRET The first genetically encoded...

  16. CRISPR Plasmids - C. elegans

    Type
    Collection
    ...Cas9 plasmid Joung 46170 PU6::klp-12_sgRNA cU6 PCR (see paper) Transfection none, need Cas9 plasmid Calarco...gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter...with a wide variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance...Calarco 47943 pMB70 cU6 BsaI Transfection none, need Cas9 plasmid Boxem 47941 pMB60 T7 BsaI In vitro transcription...elegans none, need Cas9 plasmid Ristow Do you have suggestions for other plasmids that should be added to this...CRISPR plasmids that have been designed for use in C. elegans. CRISPR... CRISPR C. elegans CRISPR Plasmids: C. elegans...

  17. Our Most Popular Blog Posts from 2019

    Type
    Blog Post
    ...feature some recently deposited plasmids every quarter. The “hot plasmids” from last March were quite popular...behalf of scientists. 9. Plasmids 101: Positive and Negative Selection for Plasmid Cloning by Jennifer Tsang...changes.   1. Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection by Alyssa...past year: CRISPR, optogenetics, chemogenetics, plasmids, fluorescent proteins, careers, and more. These...s the top 10 blog posts from the year. 10. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent... about mNeonGreen, mTFP1, and mWasabi and find plasmids containing these fluorescent proteins at Addgene... Read other Addgene news Find the most recent plasmid blog posts Find the most recent CRISPR blog posts...

  18. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...Addgene blog:  Plasmids 101: What is a plasmid? Plasmids 101: Origin of ReplicationPlasmids 101: PCR...process known as plasmid multimerization. Plasmid Multimerization Early studies on plasmid topology recorded...ligate the plasmid, and re-transform the ligated plasmid to recover the monomeric form of the plasmid. Have... monomer of Plasmid 12259: pMD2.G (depicted on the left) is ~5.8 kb in length. The plasmid can exist on... have been utilizing full plasmid sequencing services to verify the plasmids, including long-read sequencing... the sequencing results for their plasmids, they found the plasmids were much longer than expected, and...tandem. Such plasmids are known as multimers, and they are the result of multiple plasmid copies combining...

  19. Antibody Production

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    Collection
    ... low levels of co-purified plasmid from transfection. This residual plasmid is extracted and amplified...vectors, human embryonic kidney cells are transfected with the plasmid(s) encoding the heavy and light chains...compared to an untransfected control and the protein size is confirmed. Residual Plasmid Sanger Sequencing...the Sanger method, and aligned with the parental plasmid sequence to confirm the antibody’s identity. For... The fluorescence pattern is compared to an untransfected control. Immunohistochemistry (IHC) Tissue sections...

  20. Luciferase Plasmid Collection

    Type
    Collection
    ...Luciferase Plasmids Luciferase Plasmid Collection Empty Backbones Expression Constructs Reporter Constructs...otherwise noted, these plasmids are meant for expression in mammalian systems. ID Plasmid Luciferase Type(s...preparations of select luciferase expression plasmids. ID Plasmid Luciferase Type Promoter Description PI ...AAV-CRISPR knockout plasmid. Renilla luciferase is used to monitor viral transfection Feng Zhang 21471 pLenti...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...of luciferase plasmids for gene expression assays and bioluminescent reporters. Plasmid... Plasmid Collections Luciferase...
Showing: 41 - 60 of 134 results