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We narrowed to 509 results for: plasmid dna

Showing: 301 - 320 of 509 results

  1. 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...

  2. 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 ...

  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. 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...

  5. 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 (...

  6. 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-...

  7. 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...

  8. 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...

  9. Cultivating Community Science at BosLab

    Type
    Blog Post
    ...Kotton Learn How Plasmid Sharing Speeds Science Resources at Addgene.org Order Plasmids from Addgene! ...to engineer microbes. We’ve ordered a few yeast plasmids from Addgene and are eager to do our first yeast...community to collect various soil samples, extract DNA, sequence it, and analyze the data to characterize...

  10. 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." ...

  11. 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...

  12. Antibodies 101: Flow Cytometry

    Type
    Blog Post
    ... viability controls such as DNA binding dyes and amine reactive dyes. DNA binding dyes, such as propidium...membranes as is the case with dead or dying cells. DNA binding dyes are not compatible with protocols that...amine reactive dye, such as LIVE/DEAD stain. Like DNA binding dyes, amine reactive dyes can only pass through...Visit the Fluorescent Protein Guide Find antibody plasmids for your experiments ...

  13. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    ...in a 1-plasmid system, in which Cas9 is included on the gRNA-containing plasmid, or a 2-plasmid system...contain thousands of gRNA plasmids, discerned only by a unique barcode on each plasmid. As such, sequencing...gRNA generates a double-strand break in the target DNA, which, when repaired through error-prone non-homologous...are composed of many different gRNA-containing plasmids, each with a different genomic target. The goal...sequences and ease of generating gRNA-containing plasmids. CRISPR genome wide-screening experiments commonly...other types of experiments, check out our CRISPR Plasmids and Resources Page. References McDade JR, Waxmonsky...Lentivirus Get Tips on Titering Your Lentivirus Plasmids 101: NGS Quality Control for Pooled Libraries ...

  14. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    ...subfields.  Plasmid requests correlate with paper publication The dataset includes 11,426 plasmid requests...paper has between 1–32 plasmids and a total of 2–2,590 requests. We received plasmid requests from 56 countries...Spikes in plasmid requests correspond to publications of popular papers. Figure 1: Plasmid requests...number of plasmids ordered for each paper. Rough estimation of COVID-19 effect on plasmid requests ...majority of our plasmid orders come from the U.S. and Europe, we see a corresponding dip in plasmid request activity...of how plasmid orders were affected by the COVID-19 pandemic, we compared the number of plasmid orders...Learn more about plasmid sharing during COVID-19 Resources on Addgene.org Find plasmids from David Liu's...

  15. Designer PUF Proteins for Any RNA Target

    Type
    Blog Post
    ...transcription factors – to specific sequences of DNA has become commonplace. This ability has opened up...and improve, Abil says they will make those new plasmids available through Addgene, and they hope others...

  16. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...Addgene.org Find plasmids from the Bethune Lab Find Plasmids from the Roux Lab Find Plasmids for FRET ...Learn about plasmids for in vivo biotinylation of bacterial fusions proteins Study Protein-DNA Interactions...BirA* fusion proteins are expressed in cells via plasmids or by the creation of stable cell lines and these...Interested in using Split-BioID with your research? Plasmids are available from Addgene! References Schopp... Central PMCID: 5467174. Find the Split-BioID Plasmids Here! Munter, S.D., Görnemann, J., Derua, R.,...site than in Schopp et al. Find these Split-BioID Plasmids Here!   Burke, B., Kim, D.I., Roux, K.J., & Raida...PubMed Central PMCID: 3308701. Find the BioID Plasmids Here! Macneill, S., & Varnaitė, R. (2016). Meet...

  17. Open Enzyme Collection

    Type
    Collection
    ...Synthetic Biology Plasmids BIOFAB Plasmid Set Drew Endy Lab Plasmids Jennifer Molloy Lab Plasmids Blog: Golden...assembly. Plasmids Explore the collection and order individual plasmids via the links below: DNA Ligases... ID Plasmid Gene/insert 165503 pOpen-HSlig4 DNA Ligase IV 165514 pOpen-9N7lig 9 Degrees North DNA Ligase...pOpen-Pfulig Pfu DNA Ligase 165535 pOpen-THEAQlig Taq DNA Ligase 165554 pOpen-T4gene30 T4-DNA Ligase 165564...pOpen-ECOligA E. coli DNA Ligase 165569 pOpen-Tthlig Tth DNA Ligase 165574 pOpen-HSlig3 DNA Ligase III Return... to top DNA Polymerases Addgene ID Plasmid Gene/insert 165500 pOpen-9N7polA 9 degrees N-7 DNA polymerase...Tli DNA polymerase (exo-) 165524 pOpen-T4gene43 T4 DNA Polymerase 165532 pOpen-ECOpolA E. coli DNA Polymerase...

  18. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...for verifying plasmids like this is "plasmid fingerprinting", where you cut the plasmid into 3-8 pieces...Digest You may also like... Restriction Digest of Plasmid DNA Agarose Gel Electrophoresis Introduction Restriction... to selectively move a specific piece of DNA from one plasmid to another. A diagnostic restriction enzyme...enzymes cleave DNA at specific sequences called restrictions sites. Often, the size of the plasmid insert and...used to verify your plasmid. The goal of a diagnostic digest is to cut your plasmid into specific sized...restriction enzyme or enzymes that cut your plasmid. Many DNA analysis tools, including Addgene’s Sequence... Pipetman Pipet tips Reagents Liquid DNA aliquot of your plasmid of interest (see below for recommended...

  19. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head...Hutchinson Cancer Research Center developed a pooled plasmid library encompassing all possible amino acid mutations...anti-SARS-CoV-2 antibodies. In the library, each plasmid encodes the mutated spike protein and a barcode...head over to our CRISPR Plasmids and Resources page.  A new system uses light to deactivate photocleavable... accumulation of indels to measure time. These plasmids with different promoters driving expression of...

  20. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...consisted of three plasmids: the packaging, envelope, and transfer plasmids. The packaging plasmid carried a mutated... 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 ...genome integration. After co-transfection of these plasmids, 293T cells released transgene-containing lentiviral...recombination between the transfer and packaging plasmids does exist. To further reduce this possibility...
Showing: 301 - 320 of 509 results