We narrowed to 511 results for: plasmid dna
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TypeCollection...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...
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ReFigure: Save Scientific Figures into Dashboards and Share Your Insights
TypeBlog Post...whether Natronobacterium gregoryi Argonaute has any DNA editing functionality Collecting preclinical and ...Sharing Resources at Addgene.org Deposit Your Plasmids with Addgene Find Molecular Biology Protocols ... -
Multiple Plasmids at a Low Price: Kits from Addgene
TypeBlog 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 ... -
CRISPR Plasmids and Resources
TypeCollection...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... -
AAV Production in HEK293 Cells
TypeProtocol...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... -
Antibodies 101: Producing Recombinant Antibodies
TypeBlog 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... -
Cpf1 Update: Comparison to Cas9 and NgAgo
TypeBlog 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 ... -
CRISPR 101: Validating Your Genome Edit
TypeBlog 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 ... -
General Transfection
TypeProtocol...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 ... -
CRISPR Plasmids - Plants
TypeCollection...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...paired with a wide variety of Cas-containing plasmids. ID Plasmid Promoter Cloning Enzyme Co-expressed Cas9...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the DSB and a...expression plasmids that have been designed for use in plants. CRISPR... -
Synthetic promoter AAVs for cell-type specific expression in retinal cells
TypeBlog 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... -
Top Requested Lentivirus and AAV of 2016
TypeBlog 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... -
Viral Vectors 101: Biosensors
TypeBlog 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 ... -
Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity
TypeBlog 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 ... -
CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt
TypeBlog Post...Chris Voigt for speaking with us! Find the Plasmids: Plasmids for multi-input CRISPR/Cas9 genetic circuits...orthogonality. You can create an almost limitless number of DNA binding proteins with Cas9 guided by different promoters... -
Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens
TypeBlog 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... -
Transfection for Recombinant Antibodies
TypeProtocol...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... -
Protocol - How to Purify DNA from an Agarose Gel
TypeProtocol...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... -
Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping
TypeBlog 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 ... -
CRISPR Plasmids - Drosophila
TypeCollection...CRISPR Plasmids: Drosophila Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime...gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert Promoter...with a wide variety of Cas-containing plasmids. ID gRNA Plasmid Promoter Cloning Enzyme(s) Delivery Resistance... Cas9 plasmid Bullock and Port 51026 U6-BbsI-crRNA dU6 BbsI Transfection none, need Cas9 plasmid O'Connor-Giles...suggestions for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail...changes, researchers use ssDNA or dsDNA repair templates with homology to the DNA flanking the DSB and a...CRISPR plasmids that have been designed for use in Drosophila and other insects. CRISPR...