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We narrowed to 508 results for: dna plasmids

Showing: 301 - 320 of 508 results

  1. What the HEK?

    Type
    Blog Post
    ...transfect the ‘helper’ plasmids (viral packaging machinery) along with the viral plasmid containing the cargo...Netherlands. Transformation of HEK cells with adenovirus 5 DNA was postdoc Frank Graham’s 293rd experiment, hence...T-antigen. This allows them to replicate viral plasmids with the SV40 origin of replication. To generate...

  2. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ... Protocols DNA Ligation DNA Ligation You may also like... Plasmid Cloning by Restriction...and insert DNA to use. Combine the following in a PCR or Eppendorf tube: Vector DNA Insert DNA Ligase Buffer...How to Ligate Plasmid DNA Protocols...Restriction Enzyme Digest Restriction Digest of Plasmid DNA Bacterial Transformation Background Information...the construction of a recombinant plasmid is connecting the insert DNA (gene or fragment of interest) into...backbone of the two DNA fragments. This reaction, called ligation, is performed by the T4 DNA ligase enzyme....ligation, the insert DNA is physically attached to the backbone and the complete plasmid can be transformed...

  3. Protocol - Bacterial Transformation

    Type
    Protocol
    ...also like... Plasmid Cloning by Restriction Enzyme Digest Restriction Digest of Plasmid DNA DNA Ligation ... will help to maintain the plasmid without rearrangement of the plasmid DNA. Additionally, specific treatments...tube of plasmid DNA and just need to transform bacteria so that you can grow up more of the plasmid) you ... which foreign DNA is introduced into a cell. Transformation of bacteria with plasmids is important not...both storing and replicating plasmids. Because of this, nearly all plasmids (even those designed for mammalian...the least expensive) are fine for transforming plasmid DNA for the purposes of storage and amplification...transforming with very small amounts of DNA or if you're multiple plasmids at once. To save money, many labs...

  4. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    ... best bet is the Plasmid Map: We auto-generate a plasmid map based off of the plasmid sequence and feature...colleagues. Top of the plasmid page The ‘breadcrumb’ trail at the top of any plasmid page gives the hierarchy... to the individual plasmid page. The depositing lab page contains all the plasmids from the same lab while...contains all the plasmids deposited from the same publication. Next up, we have the plasmid name: This is...other plasmids with similar names in the collection, but you can use this name to find this plasmid from...A quicker way to find this plasmid through search is... The Addgene plasmid number: This is the unique... given to every plasmid in our catalog - if you know the catalog number of the plasmid you are searching...

  5. CRISPR Plasmids - Mammalian Expression

    Type
    Collection
    ...CRISPR Plasmids: Mammalian Expression The following CRISPR plasmids have been designed for use in mammalian...NHEJ. ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Browse CRISPR Plasmids By Function...specific base editor that can alter DNA bases without inducing a DNA break. Base editors convert C->T (...gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter...gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene/Insert Promoter... purify a region of genomic DNA and its associated proteins, RNA, and DNA. The enCHIP system uses an anti-FLAG...variety of Cas-containing plasmids. Do you have suggestions for other plasmids that should be added to ...

  6. Gibson Assembly Protocol

    Type
    Protocol
    ... your plasmid, think about what DNA segments you will need to join to create your final plasmid. Adjacent... single-strand DNA 3’ overhangs by chewing back from the DNA 5’ end. Complementary DNA fragments can subsequently...reaction. Can combine many DNA fragments at once. Procedure Design your plasmid and order primers (see figure...making a small change in a plasmid (such as point mutations). Generate DNA segments by PCR. Run PCR product...High-Fidelity DNA Polymerase - incorporates nucleotides to “fill in” the gaps in the annealed DNA fragments...fragments. Taq DNA Ligase - covalently joins the annealed complementary DNA fragments, removing any nicks and...creating a contiguous DNA fragment. Pro-Tip Add Extreme Thermostable Single-Stranded DNA-Binding protein (...

  7. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ... multiple plasmids. Second generation packaging systems use 3 plasmids; an envelope plasmid that usually...encodes VSV-G, a packaging plasmid containing structural genes and a transfer plasmid carrying the transfer...Rev gene from the packaging plasmid and incorporating it into a fourth plasmid. In addition, the promoter..., they also require a fourth plasmid. The requirement for additional DNA can impact the transfection efficiency...and packaging plasmids in order to obtain the best titers. Requiring an additional plasmid for transfection...By dividing the viral elements across multiple plasmids, the risk of recombination and consequently, the...thereby affect the titer of the virus. Additional plasmids will also affect experiment optimization. Before...

  8. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ...transfer plasmid), the envelope (Env) gene on a separate plasmid (envelope plasmid), and the polymerase ...different plasmids. For a retroviral system this looks like the genetic cargo in one plasmid (the transfer...Gag structural protein on yet a third plasmid (packaging plasmid). Viral production then proceeds by transfection... transfection of all three plasmids into a producer cell line, such as HEK-293T, production of viral particles... and Pol genes are not encoded in the transfer plasmid, the component that physically integrates into ... retroviral packaging components onto distinct plasmids for safety.    Viral cargo-specific safety...they physically deposit their cargo into the host’s DNA. These viruses also have a propensity to integrate...

  9. A Novice's Perspective on Neuroscience - SfN2017

    Type
    Blog Post
    ... If there’s a plasmid or AAV tool that you can’t find, you can always suggest a plasmid to us, request...want to get into the field. Find Optogenetics Plasmids for Use in Neuroscience Research Neuroscience ...images of brain regions, or sequences of microbial DNA they all seem to require some mathematical know-how...400 who signed up for our newsletter and a free plasmid) were very excited about our new viral services...Service Find Retrograde AAV Browse Optogenetics Plasmids ...

  10. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    ...successful in your organism or cell type? You can use DNA sequencing or other molecular cloning techniques ...and to make the desired genome edit:  A donor plasmid that contains a puromycin resistance gene linked...the gRNA used when transfecting your cells.  A plasmid that contains Cas9. The Madison lab also created...in the reporter gene). A control RFP expression plasmid.  With this set up, all successfully transfected... and repaired via HDR using the supplied donor plasmid will express GFP. If there’s GFP expression, presumably...just your desired genomic edit.  Find SRIRACCHA plasmids at Addgene GEMCherry2, an improved monomeric fluorescent...embryonic stem cells.  Find GEmCherry2 reporter plasmids at Addgene Overall SRIRACHA and GEmCherry2 reporter...

  11. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...region of a gene (cDNA) can be done by either plasmid or virus. A plasmid carrying your cDNA can be targeted...resources on the Addgene blog Plasmid 101: Inducible Promoters Plasmid 101: Repressible Promoters CRISPR...knock out. Looking for some permanent knock out plasmids? Check out our Cre vector and Murine LoxP vector...will inhibit transcription instead of introducing a DNA double strand break. Transient introduction of CRISPRi...of protein expression. Addgene has a dCas9 KRAB plasmid for transient knockouts!  Turn it on Turn it on...to integrate your choice cDNA. Both lentivirus and retrovirus can package cDNAs for genomic delivery and...to choose an appropriate promoter to express your cDNA, depending on the level of expression you would ...

  12. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

    Type
    Blog Post
    ...study of non-coding regulatory elements. Regulatory DNA function is typically inferred based on correlative...procedures. The video details how to transfect CRISPR plasmids into MEL cells by electroporation, sort your transfected... PubMed. More CRISPR Refereces: Browse CRISPR Plasmids CRISPR Information at Addgene More CRISPR-...

  13. Cre-Lox and Other Site-Specific Recombinases

    Type
    Collection
    ...Site-Specific Recombinases Cre Plasmids Flp Plasmids Dre Plasmids Other Recombinase Plasmids Recombinase-Dependent... cleave all four DNA strands at once. They can mediate the integration of a large DNA payload containing...same DNA strand in opposite directions, recombination results in an inversion and the region of DNA between... circular piece of DNA (and is not maintained). C) If the sites are on separate DNA molecules (in trans...recombination of the DNA between target sites. Codon optimization: Modifying the DNA sequence of the recombinase... vectors. Highlighted Plasmids Browse the tables below to find popular plasmids expressing your favorite...tamoxifen”, or “light”): Cre Recombinase Plasmids ID Plasmid Description Vector Type Promoter PI Additional...

  14. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    ...restriction digest of plasmid DNA, and bacterial transformation. And more plasmid-related educational resources... requested plasmids. I wondered how many scientists have used the protocol for this plasmid, and if it... your protocols onto your plasmid page, scientists who request your plasmid will have easy access to your...be found in our Plasmids 101 blog series. Want to learn more about sharing your plasmids via Addgene? Check... resources, such as protocols, Plasmids 101 articles on various plasmid features, and reference tables...deposit their plasmids to the repository. These protocols show up on the related plasmid pages and can...http://www.addgene.org/ If you’re working with plasmids and you decide to share them using Addgene’s repository...

  15. CRISPR Plasmids - Yeast

    Type
    Collection
    ...cytosine base editor that can alter DNA bases without inducing a DNA break. Cytosine base editors convert...gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter...gRNA expression plasmid to target the dCas9-repressor to your specific locus. ID Plasmid Gene/Insert Promoter... purify a region of genomic DNA and its associated proteins, RNA, and DNA. The enCHIP system uses an anti-FLAG...with a wide variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Validated In...cerevisiae LEU2 none, need Cas9 plasmid Lu Do you have suggestions for other plasmids that should be added to ...changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the DSB and a...

  16. CRISPR Plasmids - Cascade-Cas3

    Type
    Collection
    ...family. After nicking the target DNA, Cas3 initiates single-strand DNA degradation. This degradation can...CRISPR-Cas3 plasmids take an all-in-one approach and include Cas3 and Cascade on one single plasmid, though...Mammalian ID Plasmid Gene/Insert Promoter PI Publication Return to top Bacteria ID Plasmid Gene/Insert ...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...Find plasmids for CRISPR-Cas and Cascade for large genomic deletions. CRISPR... CRISPR Cas3 CRISPR Plasmids: Cascade-Cas3...

  17. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    ...transfection of single plasmids vs pooled plasmids; for multiple edits, single plasmids are generally more...transfected on a single plasmid. This is versus 2.4% of cells transfected with a pool of plasmids that contained...sites. This eliminates bulk from crRNA expression plasmids resulting in smaller, more streamlined vectors...contained one crRNA per plasmid plus a Cpf1 expression vector.  See the graph in figure 2 for a comparison of...  Lentiviruses carry a (+) strand RNA copy of the DNA sequence, which is a suitable substrate for Cpf1....your research? Check out the Cpf1 multiplexing plasmids available from Addgene. If you have any questions... Our CRISPR Multiplexing Resources Find CRISPR Plasmids for Your Research Catch up on Your CRISPR Background...

  18. CRISPR Library Amplification

    Type
    Protocol
    ...pooled library DNA (100 ng plasmid DNA per 25 µL electrocompetent cells) Less library DNA can be used but...multiplexed experiments. These pooled plasmids differ by only a short region of DNA: guide RNAs targeting different...differences, and plasmid recombination can all impact the representation of individual plasmids in the pooled... ). Lentiviral plasmids can recombine between their LTRs resulting in a smaller plasmid containing only... to ensure high efficiency uptake of plasmid library DNA. This quantity of cells is sufficient for libraries...200,000 individual plasmids. For larger libraries scale the number of cells and DNA accordingly. Pro-Tip...enough for the purpose of a Maxiprep. Purify plasmid DNA using the Qiagen HiSpeed Maxi Kit (one conical...

  19. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ...posts , or find CRISPR plasmids for your research. Addgene has CRISPR plasmids for the following models...editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature . 533(7603):420-4....Fonfara I, Hauer M, Doudna JA, Charpentier E. 2012. A programmable dual-RNA-guided DNA endonuclease in adaptive...specifically, specialized Cas proteins snip foreign DNA into small fragments approximately 20 bp in length...length and paste them into contiguous stretches of DNA known as CRISPR arrays. Separate Cas proteins then...motif (PAM). The CRISPR complex binds to the foreign DNA and cleaves it to destroy the invader ( Figure 1 ...protospacer sequences that have homology to foreign DNA. Protospacers are separated by short palindromic ...

  20. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    ... about FLEx vectors Download our Plasmids 101 eBook Read our plasmids blog posts Resources on the Addgene...molecular biology reference Find Cre-lox plasmids at Addgene Browse plasmids at Addgene ...site-specific recombinases such as Cre or Flp that induce DNA recombination at defined recognition sites (i.e.,...Wolf, and R. J. Samulski. "Delivering transgenic DNA exceeding the carrying capacity of AAV vectors." ...
Showing: 301 - 320 of 508 results