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We narrowed to 509 results for: plasmid dna

Showing: 341 - 360 of 509 results

  1. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ...Once you've gotten to know what a plasmid is in general, it's time to take a look at some of its parts...origin of replication are necessary to replicate plasmids and make sure the cells maintains it. Here, we'll...the promoter, which is necessary for getting the plasmid express our gene of interest. This element is responsible...  The RNA polymerase(s) RNA is transcribed from DNA using an RNA polymerase (RNAP). In bacteria this .... This means that the promoter present in your plasmid backbone must to be compatible with the type of...transcript, you will also need to make sure your plasmid has a promoter suited to working in your host organism...development-specific promoters available to scientists. Plasmids are oftentimes put to therapeutic uses, and in...

  2. 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...

  3. CRISPR Guide

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    Collection
    ...purify specific regions of DNA, image DNA in live cells, and precisely edit DNA and RNA. The ease of generating...Browse Plasmids: RNA Targeting , RNA Editing Degrading DNA with Cas3 Rather than initiating DNA cleavage...-stranded DNA plasmid. Small ssDNA oligos often work well for small edits like a single-point mutation...homology with the target DNA. Once the Cas9-gRNA complex binds a putative DNA target, the seed sequence...will begin to anneal to the target DNA. If the seed and target DNA sequences match, the gRNA will continue...strands of the target DNA. This results in a double-strand break (DSB) within the target DNA, located ∼3–4 nucleotides...of target DNA. Cas9n increases target specificity. Since two nickases targeting opposite DNA strands are...

  4. CRISPR Plasmids - Bacteria

    Type
    Collection
    ...efficiency than NHEJ. ID Plasmid Gene/Insert Promoter PI Publication Browse CRISPR Plasmids By Function Genome...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...Ampicillin none, need Cas9 plasmid Qi Do you have suggestions for other plasmids that should be added to ...

  5. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ... Check out All the Plasmids from the Collin's Lab Find Synthetic Biology Plasmids for Your Next Experiment... days of lost productivity waiting for follow-up DNA oligos to arrive. We also were careful to set up ...Components." Cell (2016). PubMed PMID: 27160350. Find Plasmids from the publication at Addgene 2. Green, Alexander...Experiment Deposit Your Plasmids with Addgene ...

  6. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...measure the DNA concentration, use 1 μL) 2 μL 10x NEB T4 DNA ligase buffer 1 μL NEB T4 DNA ligase add ...transfect plasmid DNA directly into the target cells. The shRNA will be expressed, but the DNA is unlikely...μg pLKO.1 shRNA plasmid 750 ng psPAX2 packaging plasmid 250 ng pMD2.G envelope plasmid to 20 μl serum-...to vary the ratio of shRNA plasmid, packaging plasmid, and envelope plasmid to obtain the ratio that gives...shRNA. psPAX2 Packaging plasmid for producing viral particles. pMD2.G Envelope plasmid for producing viral... also be used with packaging plasmid pCMV-dR8.2 dvpr and envelope plasmid pCMV-VSVG from Robert Weinberg...#B7001S NEB buffer 2 NEB #B7002S T4 DNA ligase NEB #M0202S T4 DNA ligase buffer NEB #B0202S DH5 alpha ...

  7. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...Storage of Plasmids Recovering Plasmid DNA from Bacterial Culture Background Information Plasmids can carry...isolate your plasmid DNA from the bacterial culture by following Isolating Your Plasmid DNA . Antibiotic...copy regardless of plasmid size. See the plasmid's info page to determine if your plasmid is high or low ...resistance gene on a plasmids allows researchers to easily isolate bacteria containing that plasmid from bacteria...numbers of bacteria necessary to isolate enough plasmid DNA for experimental use. The following protocol ... low copy plasmids? The copy number refers to the number of copies of an individual plasmid within a single...number plasmids should only need to be grown for 12-16 hr on average. Certain features of a plasmid may ...

  8. A Deep Dive into BioBuilder

    Type
    Blog Post
    ...Standards Find Plasmids for Creating and Controlling Gene Networks Browse All SynBio Plasmids ...cellular extracts as a means to rapidly prototype DNA circuits and logic gates. This BioBuilder-Caltech...

  9. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...comprehensive CRISPR guide Find CRISPR plasmids and resources Find CRISPRa plasmids ...genes can be relatively laborious because multiple plasmids/viruses are needed to deliver all the protein ...construct for generating the SPH mouse. (B) AAV plasmids used to express CRE under neuron-specific promoters...individual neuronal populations.  The Yang Lab’s AAV plasmids used to express Cre under neuron-specific promoters... CD8+ cells were then isolated and their genomic DNA sequenced. Guide RNAs targeting several known and...sensitive applications such as CRISPR screening. The plasmids used by the Qi Lab for this particular application...simpler to implement in genome-wide screens over cDNA libraries. However, using CRISPRa to target individual...

  10. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...Cas9’s second HNH endonuclease domain. Cpf1 cleaves DNA in a staggered pattern and requires only one RNA ...Francisella Cpf1 (FnCpf1), they employed an E. coli plasmid depletion assay to discover FnCpf1’s PAM sequence...system." Cell (2015). PubMed PMID: 26422227. Find plasmids from this publication at Addgene. 2. Zetsche,....1 (2016): 31-34. PubMed PMID: 27918548. Find plasmids from this publication at Addgene. 3. Kim, Daesik...Biotechnol. (2016). PubMed PMID: 27272384 Find plasmids from this publication at Addgene. Kleinstiver...Biotechnol. (2016). PubMed PMID: 27347757. Find plasmids from this publication at Addgene. Makarova, Kira...PMID: 26411297. Resources at Addgene Find CRISPR plasmids at Addgene Read Addgene's CRISPR Resource Guide...

  11. Plasmid Modification by Annealed Oligo Cloning (with Protocols)

    Type
    Protocol
    ...Restriction Digest of Plasmid DNA DNA Ligation Bacterial Transformation Summary Oligo overlap cloning can...anytime you need to add a short stretch of DNA to a plasmid, such as: Changing a Multiple Cloning Site...cut out the band containing your vector DNA. Gel purify your DNA away from the agarose using a commercially...Modify your plasmid MCS or add short sequences using oligo overlap cloning. Protocols... Protocols Plasmid Modification...Modification by Annealed Oligo Cloning Plasmid Modification by Annealed Oligo Cloning You may also like... Restriction...

  13. 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 ...

  14. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...curated collections of plasmids as kits with greatly reduced costs per plasmid. We generally create kits...just the tool you need. The plasmids in this portion of the BIOFAB Plasmid Set contain terminators of ...populations. This kit consists of a 48 plasmid adapter library, with each plasmid carrying a unique barcoded Illumina-M13F...researcher at a nonprofit institution, you can order plasmids covering a wide range of fields from Addgene. ...kits to allow you to easily use a set of plasmids together on a tight budget. To help popularize our ...order from the repository: FX Cloning Kit BIOFAB Plasmid Set pCri System Cell Free Protein Expression Kit... production (a variety of pExpression vectors) plasmids. The type IIS restriction sites used in these ...

  15. Cre-Lox and Other Site-Specific Recombinases

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    Collection
    ...Site-Specific Recombinases Cre Plasmids Dre Plasmids Flp Plasmids Other Recombinase Plasmids Recombinase-Dependent...Fluorescent Biosensors Chemogenetics Plasmid Collection Optogenetics Plasmid Collection Monosynaptic Neuronal... 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...

  16. 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...

  17. 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... the tables below for CRISPR Cascade-Cas3 plasmids. Plasmids are available for expression in mammalian...plants. Mammalian ID Plasmid Gene/Insert Promoter PI Publication Bacteria ID Plasmid Gene/Insert Promoter...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...

  18. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ... great collection of ready-to-use rAbs and rAb plasmids!). Since recombinant antibodies conveniently allow...production, reliable expression, and easy distribution as DNA (Trimmer, 2020), you may be interested in using them...antibodies can be produced through a single or dual-plasmid transfection, depending on whether the heavy and...and light chain genes are contained to one plasmid or if they’ve been separated onto two. You’ll first...first transfect your cells with your plasmid(s), and then wait 7-14 days (in mammalian cells) to collect ...on the Addgene blog Introduction to Antibodies Plasmid-based Recombinant Antibodies Polyclonal Antibodies...

  19. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    ...distant cousins of well-known Cas9. Cpf1 cleaves DNA in a staggered pattern and requires only one RNA ... is well suited for genome editing.  Find Cpf1 Plasmids at Addgene   Image attribution: Zetsche...Biotechnol. 2016 Jun 6. PubMed PMID: 27272384 Find plasmids from this publication at Addgene. 2. Kleinstiver...Biotechnology (2016). PubMed PMID: 27347757. Find plasmids from this publication at Addgene. 3. Zetsche ....cell.2015.09.038. PubMed PMID: 26422227 Find plasmids from this publication at Addgene. 4. Makarova...Our CRISPR Guide Pages Check Out MpAgo from the Doudna Lab Find Validated gRNAs ...

  20. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ... uses a 3rd sequencing trace file from the donor DNA. This method can precisely estimate editing frequency...clones. The drawback is that you must generate a DNA molecule with homology to your genome edit to mimic...Tools Read other CRISPR blog posts Find CRISPR Plasmids at Addgene  ...
Showing: 341 - 360 of 509 results