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We narrowed to 509 results for: plasmid dna

Showing: 361 - 380 of 509 results

  1. AAV Production in HEK293 Cells

    Type
    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...

  2. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    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...

  3. Viral Vectors 101: Biosensors

    Type
    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 ...

  4. Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

    Type
    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 ...

  5. Top Requested Lentivirus and AAV of 2016

    Type
    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...

  7. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    ...Konermann, the first generation of CRISPR-Cas9 plasmids were not designed with gene activation in mind...tetraloop and stem-loop 2; 3. The MS2-P65-HSF1 plasmid which expresses the activation helper protein. ...structure of Cas9 in complex with guide RNA and target DNA. Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata...

  8. 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 ...

  9. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    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...streptavidin and separated via flow cytometry.  Find plasmids for EXCELL here! Since BioID was developed in ...

  10. General Transfection

    Type
    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 ...

  11. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...Tet-Off system.  For more information on Addgene Tet plasmids, see our Tetracycline resource page. Cumate Switch...binding to the lexA operator (lexAop). Fusing the LexA DNA-binding domain (DBD) to GAL4 or VP16 activation domain...20697123. PubMed Central PMCID: PMC2942869. Find plasmids from this publication at Addgene. del Valle Rodriguez...20434990. PubMed Central PMCID: PMC2883883. Find plasmids from this publication at Addgene. Wei, Xing, ...22406855. PubMed Central PMCID: PMC3846601. Find plasmids from this publication at Addgene. Riabinina, ...25581800. PubMed Central PMCID: PMC4344399. Find plasmids from this publication at Addgene. Riabinina O...Hill, C. et al. A LexAop > UAS > QUAS trimeric plasmid to generate inducible and interconvertible Drosophila...

  12. TALEN Guide

    Type
    Collection
    ... “multiple DNA fragments in an ordered fashion in a single reaction.” Using these plasmids, which are ...Guide to TALEN technology and using TALEN plasmids TALENs...Technologies You may also like... TALEN Kits CRISPR Plasmids Zinc Finger Consortium Originally published in...Imagine being able to manipulate a specific region of DNA in the genome of your favorite species almost as ...biological disciplines are coming closer to this DNA targeting ideal. Figure 1: (A) Representation of ...enzymes and other useful protein domains to a specific DNA sequence ( Figure 1A ). Scientists have identified...expect that assembling relatively small, repeat-laden DNA regions into a single construct would be technically...

  13. CRISPR Plasmids - CRISPR Transposases (CAST)

    Type
    Collection
    ...Find plasmids for CRISPR-associated transposases (CASTs) for insertion of donor DNA....CRISPR Plasmids: CRISPR Transposases (CASTs) Browse CRISPR Plasmids By Function Genome Editing Cut Base...precise DNA-targeting of CRISPR-Cas systems, allowing for the integration of large fragments of DNA at specific...search the tables below for CRISPR Transposase plasmids. Plasmids are available for expression in mammalian...Mammalian ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...guide RNA. The transposase can then integrate donor DNA. Editing by CASTs is not considered "scarless", as...

  14. Transfection for Recombinant Antibodies

    Type
    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 Molecular Biology...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...

  15. Protocol - How to Purify DNA from an Agarose Gel

    Type
    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...

  16. Genomic Deletions in Mammalian Cell Lines

    Type
    Collection
    ...usage of the pSpCas9(BB) plasmid (pX330) (Addgene plasmid ID 42230). This plasmid allows for the simultaneous...utilized, such as pX458 (Addgene plasmid ID 48138) or pX459 (Addgene plasmid ID 48139), which include GFP ...pellets in 50 μl of DNA extraction solution. NOTE: Generally ~100,000 cells are used for DNA extraction, although...recommend an efficient method for DNA extraction, any method for genomic DNA isolation may be utilized to ...evaluate clones at the DNA as well as RNA and/or protein levels. To evaluate the DNA, amplify deletion bands... with a PCR cloning kit) into a plasmid vector. Transform the plasmid into DH5α E. coli cells and plate...Physiologica Scandinavica . 2003. PubMed . Purifying plasmid DNA from bacterial colonies using the Qiagen Miniprep...

  17. CRISPR 101: Cas9 Nickase Design and Homology Directed Repair

    Type
    Blog Post
    ...In this post, we’ll summarize how IDT (Integrated DNA Technologies) first demonstrated how CRISPR nickases...Check out CRISPR Topic Page Find CRISPR Nickase Plasmids Browse the CRISPR Guide ...insertions (long ssDNA; IDT Megamer® ssDNA Fragments) Test both bottom and top strand ssDNA donors (if possible...longer insertion (mCherry) using IDT Megamer long ssDNA with 100 nt homology arms. Schubert and Yan next...

  18. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...popular split-FP plasmids from our catalog to help you find the right split-FP plasmid for your experiment...below, and click the Article link to find all the plasmids from the associated paper:   Color Base FP...Constant rate of p53 tetramerization in response to DNA damage controls the p53 response. Gaglia G, Lahav...and experimental design. We hope you find these plasmids useful. Good luck, and happy experimenting! References...

  19. CRISPR Plasmids and Resources

    Type
    Collection
    ...select CRISPR lentiviral plasmids. CRISPR Plasmid Kits : Find groups of plasmids that are used together ...gRNA plasmids to target specific genes. Browse Plasmids by Model Organism Bacteria CRISPR plasmids designed...resources or browse plasmids below by function. Addgene's CRISPR collections feature plasmids for a variety ...information into a specific DNA site. Base Edit Base editors precisely edit DNA without promoting DSB formation.... elegans CRISPR plasmids designed for use in C. elegans . Drosophila CRISPR plasmids designed for use...Mammalian CRISPR plasmids designed for use in mammalian systems. Parasites CRISPR plasmids designed for use...parasites. Plant CRISPR plasmids designed for use in plants. Yeast CRISPR plasmids designed for use in yeast...

  20. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ... brightness and high degree of overlap. Dyes for DNA/RNA staining should have high affinity and specificity...specifically to the AT-rich regions of double-stranded DNA and exhibits a blue-fluorescent light when excited...cell and protein analysis products. You can find plasmids deposited by Promega here.    References and ...More resources from Addgene Fluorescent protein plasmids and resourcesA practical approach to choosing ...
Showing: 361 - 380 of 509 results