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We narrowed to 508 results for: dna plasmids

Showing: 341 - 360 of 508 results

  1. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...reconstituted after integration; and a sequence of stuffer DNA after the gRNA target site. Donor B contains: 400...Since this approach is dependent on double-strand DNA break repair, if non-homologous end joining (NHEJ...2013) Oversized AAV Transductifon Is Mediated via a DNA-PKcs-independent, Rad51C-dependent Repair Pathway...therapy Learn about the parts of an AAV transfer plasmid Resources on Addgene.org: Check out the Addgene...

  2. CRISPR Plasmids - Double-Strand Break (Cut)

    Type
    Collection
    ...CRISPR plasmids that create double strand breaks in DNA....genomic changes, researchers use DNA repair templates with homology to the DNA flanking the DSB and a specific...Bacteria ID Plasmid Gene/Insert Promoter PI Publication Return to top Drosophila ID Plasmid Gene/Insert...elegans ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Return to top Yeast ID Plasmid Gene... ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Return to top Xenopus ID Plasmid Gene...selection of CRISPR plasmids and resources. Find more CRISPR functions along with plasmids categorized by ...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...

  3. CRISPR Plasmids - Drosophila

    Type
    Collection
    ...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... Cas9 plasmid Bullock and Port 51026 U6-BbsI-crRNA dU6 BbsI Transfection none, need Cas9 plasmid O'Connor-Giles...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the DSB and a...CRISPR plasmids that have been designed for use in Drosophila and other insects. CRISPR...

  4. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...You may also like... Plasmid Cloning by PCR Restriction Digest of Plasmid DNA DNA Ligation Background ...stranded DNA template strands. The DNA polymerase is also stable enough to now bind to the primer DNA sequence...which would denature DNA polymerases from other species. In addition to the template DNA and the Taq polymerase...several micrograms of target DNA from just a few nanograms of template DNA through several cycles of denaturation...this step enables the DNA polymerase to synthesize and elongate the new target DNA strand accurately and... are using the same template DNA for all your reactions, the Template DNA can be added to the master mix...the number of nanomoles of DNA times 10. For example, if your lyophilized DNA is 38.5nm, add 385µl of water...

  5. Lentivirus Production

    Type
    Protocol
    ...Transfection grade DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and Transfer plasmid encoding your...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ...transfection plasmids into OptiPro SFM: Reagent Amount per 10 cm dish* Packaging Plasmid ( psPAX2 ) 1.3...1.3 pmol Envelope Plasmid ( pMD2.G ) 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume...volume 500 μL *Plasmid concentrations and ratios should be optimized for each transfer plasmid. Pro-Tip Endotoxins...dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would be 83.4 μL of 1 mg/mL PEI in 416.6 μL of OptiPro...the diluted DNA mixture. Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate...

  6. 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...enhancers in mammalian cells. These plasmids allow for cloning DNA fragments encoding TAL effector repeat...TALE Toolbox 49622 - 49647 24 plasmids Thoru Pederson TALEColor plasmids: destination vectors for use ... repetitive DNA sequences in human chromosomes Do you have suggestions for other plasmids that should ...module plasmids and improve the success rate of Golden Gate assembly. Destination vectors, pcDNA-TAL-NC2... TALE plasmids. Genome...

  7. Protocol - How to Streak a Plate

    Type
    Protocol
    ...for DNA purification will minimize the chance of having a mixture of plasmids in your purified DNA. This...Agar Plates Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture Introduction If you have... stab culture of bacteria and need to purify plasmid DNA from it, you will want to isolate an individual...single colonies, you can proceed to Recovering Plasmid DNA or use the individual colonies for other experiments...antibiotic. Label the bottom of the plate with the plasmid name and the date. It is also a good idea to add...overnight (12-18 hours) at 37 °C. Pro-Tip Some plasmids or bacteria need to be grown at 30 °C instead ... 37 °C. This is often true for large unstable plasmids, which sometimes recombine at 37 °C. Be sure to...

  8. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...taken up the plasmid. This can serve as an alternative to antibiotic selection. Plasmids of this type .... Scientists can easily utilize GFP-containing plasmids as a means to many functional ends. We’ve listed...each protein or domain being tested. Find FRET plasmids here. Split EGFP: An alternative to FRET, split... marking/selection: Expression constructs like plasmids often include GFP as a marker to help identify...nude mice in cancer studies, act as a reporter for DNA double strand break repair, and to label pathogenic...techniques in fluorescent microscopy. Read More: All Plasmids 101 Articles Addgene's Fluorescent Protein Guide... plasmids 101...

  10. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...sequence in pEcgRNA. Next both plasmids are transformed in E. coli with the donor DNA for editing to take place...beneficial for plasmid clearance. It is more convenient to update the spacer(20nt) on pEcgRNA plasmid as only.... pCutamp cures the plasmids by targeting the ampR promoter in all three plasmids. pCutamp is then removed... 13 plasmids with 14 rounds of transformation in 30 days. Figure 2: (A) MUCICAT plasmids for ... original pCas/pTargetF system consists of one plasmid that expresses the S. pyogenes Cas9, a temperature-sensitive...the pTargetF construct and then introduce both plasmids into the cell. The edit in the chromosome can ...the pMB1 replicon, resulting in cutting of the plasmid. At this point, subsequent introductions of pTargetF...

  11. Video Library

    Type
    Protocol
    ...Storing and Handling Addgene Plasmids What to do after you receive your plasmids from Addgene Instructions...Protocol Purifying DNA from an Agarose Gel Gel purification allows you to isolate and purify DNA fragments based...electrophoresis, you can cut DNA bands out of the agarose gel and purify the DNA samples. This is a commonly...Protocol Diagnostic Restriction Digest Verify a plasmid using restriction enzymes Diagnostic Restriction...Instructions for Storing and Handling Plasmids Genomic Deletions with CRISPR Generate genomic deletions in mammalian...Transformations We walk you through a standard plasmid transformation protocol and offer some tips and...the standard laboratory procedure for separating DNA by size for visualization and purification. Agarose...

  12. Celebrating 15 Years of Scientific Sharing

    Type
    Blog Post
    ...the challenges of DNA distribution to ensure scientists everywhere can access plasmids despite the evolving... that is something to recognize! Distributing plasmids, viral service, and more For the first time ...to worry about succeeding at our core mission. Plasmid distribution is amazingly still growing at >9% ...Addgene Resources on the Addgene website Find plasmids for your research  Learn about our viral service...

  13. CRISPR Plasmids - Base Edit

    Type
    Collection
    ...CRISPR plasmids for precisely editing DNA bases without inducing a DNA break....Bacteria ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Return to top Plant ID Plasmid Gene...selection of CRISPR plasmids and resources. Find more CRISPR functions along with plasmids categorized by ...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...CRISPR Plasmids: Base Edit Mammalian Bacteria Plant Yeast Zebrafish Resources Two classes of base editors ..., creating an A to G (or T to C) change. Adenine DNA deaminases do not exist in nature, but have been ...

  14. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...random) integration into the genome, and its large DNA packaging capacity is well suited for larger transgenes...AAV. Its main limitation is the small size of its DNA cargo - 4.8 kb at most - which means that a gene ...in mammals Resources at Addgene.org Find AAV plasmids Find ready-to-use AAV viral preps Browse all viral...

  15. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...episomal expression. Episomes are circular pieces of DNA that reside outside a host cell’s genome. In this... The most obvious workaround is to use a multi-plasmid system, where you have one AAV vector dedicated...levels of AAV vector integration into CRISPR-induced DNA breaks. Nature Communications, 10(1). https://doi.org...

  16. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...derivatives. Unmodified Cas9 causes double stranded DNA breaks that commonly result in a complete loss of...These mutations result in an inactive RNA-guided DNA binding protein (dCas9). dCas9 is then tethered to... CRISPR Resources on Addgene.org Find CRISPR plasmids and resources here Find pooled libraries available...viability genes as they are subject to an increase in dsDNA breaks that can cause cell death. Thus, CRISPR inhibition...mitigating toxicity induced by high copy number dsDNA cutting. A Dolcetto library containing only 3 sgRNAs...

  17. The Challenges of Cell Culture

    Type
    Blog Post
    ...good that it is used as a forensic technique (aka DNA fingerprinting) that can place suspects at the scene...range of testing, from animal doping to forensics, DNA sequencing to drug testing. You can find Nick on ...Protein Localization Resources on Addgene.org Find Plasmids for Mammalian Cells ...

  18. FlipGFP, a novel fluorescence protease reporter to study apoptosis

    Type
    Blog Post
    ... external signals such as developmental cues and DNA damage. These signals activate a cascade of caspases...these diseases and disorders. You can find the plasmids for FlipGFP and FlipCherry at Addgene. References...Download the fluorescent proteins 101 eBook Read our plasmids 101 blog series Resources on Addgene.org Browse...protein fusions Browse the fluorescent proteins plasmid kits ...

  19. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    ... How to Use the Nanobody Toolbox Find Nanobody Plasmids Structural comparison As the name suggests, ... Species Antibody Subclass Nanobody Addgene Plasmid Rabbit N/A* TP897 pTP1183 (1x cysteine) pTP955...Nanobody In Bacteria: Table 2 also has a link to the plasmid you’ll need to express your nanobody of choice ...column. bdNEDP1 protease can be generated using plasmid pDG02583. Label the Nanobodies: Purified nanobodies...Journal of Cell Biology. PubMed PMID: 29263082 Find plasmids from this publication at Addgene. 2. Bates, M...23495938. Additional Resources on the Addgene Blog DIY DNA ladders from Penn State University Use Nanobodies... on Addgene.org Find more nanobody expression plasmids here Additional Resources Looking for monoclonal...

  20. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...packaging plasmids: the pHelper plasmid containing Ad E2A, E4, and VA RNA genes; and the pRepCap plasmid containing...to be digested with DNase before undergoing titering, to remove residual plasmid DNA carried over from ...with your transfer plasmid, carrying your gene of interest, and the two helper plasmids, commonly called...called “triple plasmid transfection” (Figure 1).    Figure 1: Overview of key steps of AAV production, iodixanol...repeats (ITR). You should always have a high-quality plasmid standard and rAAV reference material of a known...blog Parts of the AAV Packaging PlasmidParts of the AAV Transfer PlasmidAAV Variables That Matter AAV protocols...
Showing: 341 - 360 of 508 results