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We narrowed to 498 results for: plasmid dna

Showing: 381 - 400 of 498 results
  1. 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...
  2. Cpf1: A New Tool for CRISPR Genome Editing

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    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...
  3. Protocol - How to Inoculate a Bacterial Culture

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    Protocol
    ...isolate your plasmid DNA from the bacterial culture by following the Isolating Your Plasmid DNA protocol....copy regardless of plasmid size. See the plasmid's info page to determine if your plasmid is high or low ...Glycerol Stocks for Long-term Storage of Plasmids Recovering Plasmid DNA from Bacterial Culture All Addgene ...resistance gene on a plasmid 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 h on average. Certain features of a plasmid may render...
  4. Multiple Plasmids at a Low Price: Kits from Addgene

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    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 ...
  5. CRISPR Plasmids - RNA Editing

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    Collection
    ...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 based on Cas13 designed to precisely edit RNA targets. CRISPR...enzymes Cas13a/C2c2 and Cas13b, target RNA rather than DNA. Fusing the catalytic domain of ADAR2(E488Q) adenosine...Browse, sort, or search the tables below for CRISPR plasmids designed for RNA editing. To learn more about ...CRISPR topics, read our CRISPR Guide . Mammalian ID Plasmid Gene/Insert PI Publication Return to top CRISPR...by organism by visiting our CRISPR plasmids page . Find a comprehensive list of CRISPR resources by visiting...
  6. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

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    Blog Post
    ... resources on the Addgene blog Plasmid 101 Protein Expression Plasmid 101 Biotinylation Other resources...vivo studies (Che and Khavari, 2017). Find APEX plasmids at Addgene! Since the development of BioID and... biotin ligase from A. aeolicus, which lacks the DNA-binding domain (233 amino acids) found in the E. ... specific locations in the cell.   Find BioID2 plasmids here APEX2: A more sensitive ascorbate peroxidase... They named this mutant APEX2.  Find the APEX2 plasmids! TurboID and minturbo: Rapid and non-toxic proximity...allowing for more precise temporal control.  Find plasmids for TurboID and miniturbo Split-BioID: A conditional...information, read the Split-BioID blog post. Find plasmids for Split-BioID RaPID: RNA-protein interaction...
  7. AAV Production in HEK293 Cells

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    Protocol
    ...of each plasmid needed to have a 1:1:1 molar ratio with 2 mg total DNA per CS5 Plasmid Plasmid Size (bp... each plasmid DNA into the bottle containing the Opti-MEM. Mix well. Add 4 mL of PEI (1:2 μg DNA to μg...conditions must be optimized. Plasmids for transfection: pHelper; pRC (Rep-Cap), plasmid expressing your gene ...(bp) DNA Concentration (μg/μL) Volume of DNA (μL) RepCap 7,265 1.00 727.6 pHelper 11,854 1.00 1,185 Transfer...Transfer Plasmid 5,842 1.00 584.2 Total bp 24,961 In total, we would like 2 mg of DNA or 2000 μg. Using...molar ratio of each plasmid: 2000 μg / 24,961 bp = 0.08 μg/bp Therefore, for each plasmid we need: Sample ... the total number of base pairs from all three plasmids, we can determine the total μg/bp we need to achieve...
  8. Antibodies 101: Producing Recombinant Antibodies

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    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...
  9. Cpf1 Update: Comparison to Cas9 and NgAgo

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    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 ...
  10. 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  ...
  11. General Transfection

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    Protocol
    ...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ...to the diluted DNA. Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate ...possible range of ratios to test: Ratio of DNA:PEI Amount of DNA (μg) Volume of 1 mg/mL PEI (μL) 1:1 18.9...protocol to transfect mammalian cells with your plasmid of interest. Protocols...25 mM chloroquine diphosphate. Dilute 18.9 µg of DNA into 500 µL of Opti-Pro SFM. Pro-Tip Endotoxins can..., the plasmid should also be propagated in an endonuclease negative E. coli strain such as NEB stable....is prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the cells 1–2 ...
  12. Transfection for Recombinant Antibodies

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    Protocol
    ...meter Reagents HEK293 cells Recombinant antibody plasmid DNA 500 mL vented flask 50 mL conical tubes Microcentrifuge...Transfer the PEI-MAX and recombinant antibody plasmid DNA sample to the biosafety cabinet and incubate ... cabinet. Add 180 µg of recombinant antibody plasmid DNA to one tube of 6 mL BCD TFX. Cap the tube and...Recombinant Antibodies You may also like... Antibody Plasmid Collection Antibodies Blog Posts Antibody Guide...suspension HEK293 cells with recombinant antibody plasmids using Polyethylenimine Max as a transfection reagent...General Considerations For antibody-expressing plasmids containing the SV40 origin of replication use ... SV40 large T antigen. For antibody-expressing plasmids containing the Epstein-Barr virus origin of replication...
  13. Protocol - How to Purify DNA from an Agarose Gel

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    Protocol
    ...like... Restriction Digest of Plasmid DNA Agarose Gel Electrophoresis DNA Ligation Background Information... Protocols Purifying DNA from an Agarose Gel Purifying DNA from an Agarose Gel You may also...which separates DNA by their length in base pairs. Following electrophoresis, you can cut DNA bands out of... minimize the risk of DNA damage, it is best to limit the UV exposure of the DNA. Therefore, it is a bad...buffer to add during the DNA isolation step. Finally, you will want to isolate the DNA from the gel. This is...intended step with the now gel purified DNA. Find more information about DNA quantification here . Tips and FAQ...Protocol for how to purify DNA from an agarose gel, including tips to help improve the resolution and...
  14. CRISPR Plasmids - Bacteria

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    Collection
    ...following CRISPR plasmids have been designed for use in bacteria. To find emtpy gRNA plasmids, visit Addgene's...Activate ID Plasmid Gene/Insert Promoter PI Publication Return to top Base Edit ID Plasmid Promoter PI... top Cut ID Plasmid Gene/Insert Promoter PI Publication Return to top Interfere ID Plasmid Gene/Insert...top Nick ID Plasmid Gene/Insert Promoter PI Publication Return to top Prime Edit 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...CRISPR plasmids that have been designed for use in bacteria. CRISPR...
  15. Synthetic promoter AAVs for cell-type specific expression in retinal cells

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    Blog Post
    ...epigenetic analysis and designed using stretches of DNA that were transcriptionally active in retinal cell... GFP marker (CatCH-GFP). The lab deposited AAV plasmids containing the active promoters at Addgene. Performance...of the retina.  Find the AAV synthetic promoter plasmids! Comparing synthetic promoters performance in ...blog posts Resources at Addgene.org Find AAV plasmids Find ready-to-use AAV viral preps Browse all viral...
  16. Top Requested Lentivirus and AAV of 2016

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    Blog Post
    ... Cas9 nuclease, which introduces double stranded DNA breaks, can be expressed from lentiCas9-Blast. Purpose...typically express Cas9 and a gRNA from the same plasmid. LentiCas9-blast virus can be used to generate ...with a single-vector system). Tips for using this plasmid Cas9 expression is typically cytotoxic, which makes...
  17. Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

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    Blog Post
    ...Visit our CRISPR plasmids and resources page Read our CRISPR guide Find CRISPR plasmids for mammalian systems...Cas9 variants with broad PAM compatibility and high DNA specificity." Nature 556.7699 (2018): 57. PubMed ...relative to Cas9. Since then, the Joung and Jennifer Doudna Labs used targeted mutagenesis to generate hyper...example is CasX, characterized by the Oakes and Doudna Labs, which is both small and potentially less ...
  18. Viral Vectors 101: Biosensors

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    Blog Post
    ...receptor could be something like an antibody, enzyme, DNA, etc. The receptors job is to generate a quantitative...genetically encoded, and luckily, we have pre-designed plasmids for these systems! Ion sensors One common biosensor... vehicle, we’ve got you covered with packaging plasmids! References and Resources References  Bhalla ...
Showing: 381 - 400 of 498 results