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  1. Viral Prep Service for Plasmids and Pooled Libraries

    Type
    Collection
    ...Virus-associated DNA Each viral vector service request will also include virus-associated DNA, which is a ...a sample of the purified DNA from the original plasmid that was used to make the virus. In order for Addgene...the right to use the original plasmid. Distribution of the original plasmid is important because it is the...Overview of Addgene's viral preparation service for plasmids and pooled libraries. Viral preparation options...produces high-quality viral vector preps from select plasmids in our collection. We know that viral vector production... production is a common step after requesting plasmids from Addgene, and by providing this service, we...options. Select from a vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need....
  2. p53 Pathway

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    Collection
    ...function in cancer. p53 Pathway Plasmids Click on a name to find available plasmids for the gene, or browse the...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...Addgene's collection of plasmids for the p53 pathway. p53 is a transcription factor and tumor suppressor...mTOR pathway Ras pathway Background p53 Pathway Plasmids p53 Gene List References Background p53 is a transcription...tumor suppressor activated by cellular stresses like DNA damage, oxidative stress, and nutrient deprivation... negative regulators, increase its stability and DNA binding activity, and allow it to bind transcriptional...of these mutations are missense mutations in the DNA-binding domain, often at one of six hotspot residues...
  3. Deep Dive: qPCR

    Type
    Blog Post
    ...single-stranded) DNA. As the temperature drops, single intensity should smoothly increase as the DNA amplicon ... are produced by the presence of double-stranded DNA, when temperatures are high, signal intensity should...dyes that are not dependent on amplification of the DNA; they are, however, affected by bubbles in wells,...LightCycler qPCR optimisation for low copy number target DNA. Journal of Immunological Methods 270 (2002) 119 ...is a smooth, single-step transition between ssDNA and dsDNA. This is not true for all amplicons; if your... probes (which you probably know as TaqMan) and dsDNA dyes (commercial name: Sybr). However, there are... Yes Yes Agarose gel or uMelt analysis dsDNA dye (Sybr)  High No No No Melt curve ...
  4. Viral Production

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    Collection
    ...the transfer plasmid, a plasmid encoding Rep and serotype-specific Cap genes, and a plasmid encoding adenoviral...Transfer Plasmid Addgene uses a rigorous barcode matching system that follows the sample from the DNA tube...cells several days later, and isolating genomic DNA. This DNA is then used as a template to amplify a portion... coli . In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits. Resources...of the viral genome and the absence of unexpected DNA contaminants. The data is also used to determine ...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...lentiviral vector, incubated for 72 h, and genomic DNA is extracted. Primers and probes targeting integrated...
  5. Protocol - How to Design Primers

    Type
    Protocol
    ...fragment of DNA. On the other hand, a long primer is used to amplify a eukaryotic genomic DNA sample. However...You may also like... Polymerase Chain Reaction Plasmid Cloning by PCR Agarose Gel Electrophoresis Primer...that are complementary to the template region of DNA. They are synthesized chemically by joining nucleotides...the primer corresponds completely to the template DNA strand so elongation can proceed. Usually a guanine...short. Short primers produce inaccurate, nonspecific DNA amplification product, and long primers result in...in a slower hybridizing rate. On average, the DNA fragment that needs to be amplified should be within...
  6. Plan Your Experiment

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    Collection
    ...Additionally, many plasmids containing validated gRNAs are now available through Addgene. These plasmids contain ...nuclease that cleaves DNA heteroduplexes, and run on an agarose gel to identify DNA fragments. PCR and restriction...amplification of targeted region from DNA, subcloning into a plasmid, and screening individual clones. PCR...particular model organism. There may not be a perfect plasmid for your specific application, and in such cases...different activators exist, including the multi-plasmid SAM system. Select Expression System To use CRISPR...to generate stable cell lines (for details, see Plasmids 101: Mammalian Vectors ) Transient or stable expression...and enrich positive cells Packaging and envelope plasmids provide the necessary components to make lentiviral...
  7. Serotype Testing AAV

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    Collection
    ...virus associated DNA (vdna), which is a sample of the purified DNA from the original plasmid that was used...AAV Vectors for Serotype Testing pAAV-CAG-GFP (Plasmid #37825) Description : Ready-to-use AAV in various... for direct injection. These AAV are made from plasmid 37825 (deposited by Edward Boyden ) and direct ...AAVrg.T 20 µL $ 140 Add to Cart pAAV-hSyn-EGFP (Plasmid #50465) Description : Ready-to-use AAV in various... for direct injection. These AAV are made from plasmid 50465 (deposited by Bryan Roth ) and direct EGFP...details, see our FAQ on What is virus associated DNA, and why do I have to order it? Don’t See What You...options. Select from a vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need....
  8. Common Injection Routes in Mice

    Type
    Blog Post
    ...Engineering Guide CRISPR Mammalian Expression plasmids Cre-Lox plasmids References Prathipati, P., Rodriguez-Aguayo...substances can be injected, including cells, drugs, DNA, viral vectors, and viruses. All researchers must...
  9. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...... Restriction Digest of Plasmid DNA Purifying DNA from an Agarose Gel DNA Ligation Introduction Gel ...length of a DNA fragment by running it on an agarose gel alongside a DNA ladder (a collection of DNA fragments...negatively charged DNA through an agarose gel matrix toward a positive electrode. Shorter DNA fragments migrate...100 mL gel). EtBr binds to the DNA and allows you to visualize the DNA under ultraviolet (UV) light. Caution...from the DNA. So if you run the gel without EtBr in the buffer you will reach a point where the DNA will ...that has UV light, visualize your DNA fragments. The fragments of DNA are usually referred to as ‘bands...or c) loading less DNA into the well. Another method for visualizing very short DNA fragments is polyacrylamide...
  10. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...creates the DNA double-strand break (DSB) at the target site, and a donor oligonucleotide or plasmid as the... Find CRISPR Plasmids for use in Mammals Check out Our CRISPR Guide Pages Find Plasmids for Mouse Targeting..., the gRNA, Cas9, and donor oligonucleotide or plasmid components are brought together and microinjected...provided (in the form of an oligonucleotide or plasmid), precise changes can be made in the genome via...mice carrying the intended mutation. With a donor plasmid, the outcome is less predictable. In our best case...
  11. KLF Research Plasmids

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    Collection
    ...of KLF plasmids and learn more about the scientists who have created and shared these plasmids....Plasmid Collections KLF Plasmids Plasmids for KLF Research KLF Depositing Labs Juan Belmonte Linzhao Cheng...information on available plasmids, references, and links. KLF1 KLF2 KLF4 KLF5 KLF6 KLF Plasmids Browse Addgene'...Addgene's collection of KLF plasmids. ID Plasmid Description Gene/Insert PI...-like factors (KLFs) are a family of at least 17 DNA binding transcriptional regulators with diverse roles...which bind to CACCC elements or GC-rich regions on DNA to mediate transcriptional activation and/or repression...shared resource on Addgene. If you have created plasmids containing KLF, we encourage you to contribute...
  12. Viral Vectors

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    Collection
    ...for select plasmids are now available through Addgene, learn more here . Browse Viral Plasmids by Type Visit... to find plasmids that can be used to engineer various types of viruses. Lentivirus Plasmids | Guide Lentiviruses...transgene expression. Overview of Viral Plasmids While plasmids are typically used to encode, propagate... this genetic information to cells. Adapting plasmid DNA for use with virus-mediated delivery has many...organism. Figure 1. Using viral plasmids to generate virus. A) Viral plasmids have been engineered such that...genomes have been adapted into a plasmid-based technology, such that plasmids can be used to create viruses...researchers can introduce plasmids to a host to generate virus. Furthermore, these plasmids can be modified to...
  13. Antibodies 101: Introduction to Antibodies

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    Blog Post
    ... be used for. Addgene’s Antibody Plasmid Collection includes plasmids that express antibodies, nanobodies... target protein. However, small mutations in the DNA of the hybridoma can occur, known as genetic drift...immunoglobulin genes, recombinant antibodies are plasmid-based. Their sequences can be optimized for specificity...protein (Wang et al., 2013). These proteins are also plasmid-derived. However, a disadvantage to ScFv is their...nanobodies, or synbodies, can be produced from plasmids, saving time and money. Nanobodies and synbodies...expression in cells that have been transfected with a plasmid encoding GFP. Actin, a common internal control ... as well as tools for developing and producing plasmid-based antibodies - and we’re always growing our...
  14. TALEN Expression Vectors

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    Collection
    ...Keith Joung Lab plasmids Daniel Voytas Lab plasmids CRISPR plasmids This page lists plasmid vectors into ...into which DNA fragments encoding TAL effector repeat arrays (assembled using the REAL, REAL-Fast or FLASH... yield TAL effector repeat arrays with identical DNA and amino acid sequences (i.e. only the details of...with the Joung Lab Individual REAL TALE Repeat Plasmids in the Joung Lab REAL Assembly TALEN Kit . Vectors...domains ( Sander et al., Nat Biotechnol. 2011 ): ID Plasmid C-terminal 0.5 TALE repeat Binds 32285 JDS70 NI...
  15. Fluorescent Proteins 101: Aptamer Fluorophores

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    Blog Post
    ...Addgene.org Find the Spinach Plasmids at Addgene Find Broccoli Plasmids at Addgene View All Fluorescent...meaning “to fit”. SELEX could be used to classify DNA aptamers as well, and over the course of the next...
  16. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ... FRET-based biosensors, and interactions between DNA and proteins. FRET, in theory, is an easy technique...used pairs to measure FRET. The table below lists plasmids that can be used to create your choice of fluorescent... fusion protein with your gene of interest: Plasmid Color Expression Description pPROEX Aqua Cyan...
  17. Zhang Lab's CRISPR Frequently Asked Questions

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    Collection
    ... single stranded DNA (ssDNA) oligo which usually work better than plasmids. For ssDNA oligo design, the... may also like... Zhang Lab CRISPR plasmids General CRISPR Plasmids Dr. Le Cong, the first author of one...easiest way is to co-transfect several plasmid together, with each plasmid having a targeting spacer cloned ...better for PCR and surveyor. I used DNA polymerase Takara Ex Taq ™ DNA Polymerase for my genomic PCR, but...consider using ssDNA oligo as donor template, it's usually more efficient than plasmid donor. Typical ...200ng to 500ng of 293FT genomic DNA as template after performing gDNA purification and have tried conditions... Genome Engineering CRISPR Zhang Lab CRISPR Plasmids Zhang Lab CRISPR FAQs Zhang Lab's CRISPR Frequently...
  18. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    ...vectors with roughly 73 million semi-random, 30 bp DNA barcodes that are integrated into target cells' genomes... ClonTracer Library at Addgene Find Lentiviral Plasmids at Addgene Check out Our Cancer Resources Page...
  19. Joung Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) Plasmids

    Type
    Collection
    ... genomic DNA sequence and a nuclease (e.g.—the Cas9 endonuclease) that cleaves the target DNA. The Joung...CRISPR-Cas/RGN expression plasmids for use in human cells CRISPR-Cas/RGN expression plasmids for use in zebrafish...Guide RNA expression plasmids for the EGFP Reporter Gene Guide RNA expression plasmids for endogenous human...RNA-Guided Nuclease (RGN) Plasmids Genome... Genome Engineering CRISPR Joung Lab CRISPR Plasmids Joung Lab CRISPR-Cas...CRISPR-Cas/RNA-Guided Nuclease (RGN) Plasmids You may also like... CRISPR Guide CRISPR Protocols gRNA Design Tools...Joung Lab CRISPR-Cas/RGN reagents: Cpf1 expression plasmids (AsCpf1 and LbCpf1) for use in mammalian cells...
Showing: 381 - 400 of 504 results