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We narrowed to 509 results for: plasmid dna

Showing: 321 - 340 of 509 results

  1. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    ...-mediated cloning CRISPR plasmids: Yeast Yeast Synthetic Biology Plasmids ...uses non-homologous end joining (NHEJ) to perform DNA double strand break repair instead of homologous ...the Addgene Blog Five Popular Model Organisms Plasmids 101: Yeast Vectors Tips for Screening with the...

  2. DNA Quantification

    Type
    Protocol
    ...Recovering Plasmid DNA from Bacterial Culture Restriction Digest of Plasmid DNA Background Information During...your DNA sample concentration and purity using Addgene’s DNA Quantification Protocol. Protocols...spectrophotometer using the solution the DNA is resuspended in, but with no DNA added. 'Blanking' measures the ... Protocols DNA Quantification...Quantification DNA Quantification You may also like... Inoculating a Liquid Bacterial Culture Recovering...important to get a quick and accurate reading of DNA concentration and purity. This can be done with a... maximally absorbs. In the case of nucleic acid (DNA and RNA), the maximal absorbance is at 260nm. Protein...

  3. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ... Some researchers have used fluorescently tagged DNA-binding proteins to label certain loci, but this ...24360272. PubMed Central PMCID: PMC3918502. Find plasmids from this publication at Addgene. 2. Ma, Hanhui...25713381. PubMed Central PMCID: PMC4364232. Find the plasmids from this publication at Addgene. 3. Ma, Hanhui...27088723. PubMed Central PMCID: PMC4864854. Find the plasmids from this publication at Addgene. 4. Kiani, Samira...26344044. PubMed Central PMCID: PMC4666719. Find plasmids from this publication at Addgene. Additional ...Resources on Asddgene.org Find Fluorescent Protein Plasmids Find Fluorescent Protein Kits Find Empty Backbones...

  4. Protocol - How to Create a Bacterial Glycerol Stock

    Type
    Protocol
    ...stocks of their plasmids. This way, when you want to make more plasmid DNA, the plasmid will already be... for long-term storage of plasmids. Although you can store your plasmid DNA at -20°C, many labs also create... Storage of Plasmids Creating Bacterial Glycerol Stocks for Long-term Storage of Plasmids You may also... like... Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture Background Information...able to start an overnight culture for plasmid DNA prep the following day. Tips and FAQ The optimal concentration...glycerol stock the same time you prepare your plasmid DNA. In the morning, when you retrieve your liquid... and growth temperature, can be found on your plasmid's information page. The next day you will be able...

  5. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ...interaction due to mismatches between the guide and the DNA target. CFD scores are then modified (“transformed...sequence, one by one, from the PAM outwards so that the DNA sequence is altered but the protein sequence they...software Resources on Addgene.org Find CRISPR plasmids and resources Download the CRISPR eBook Read our...

  6. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...reconstituted after integration; and a sequence of stuffer DNA after the gRNA target site. Donor B contains: 400...Since this approach is dependent on double-strand DNA break repair, if non-homologous end joining (NHEJ...2013) Oversized AAV Transductifon Is Mediated via a DNA-PKcs-independent, Rad51C-dependent Repair Pathway...therapy Learn about the parts of an AAV transfer plasmid Resources on Addgene.org: Check out the Addgene...

  7. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...You may also like... Plasmid Cloning by PCR Restriction Digest of Plasmid DNA DNA Ligation Background ...stranded DNA template strands. The DNA polymerase is also stable enough to now bind to the primer DNA sequence...which would denature DNA polymerases from other species. In addition to the template DNA and the Taq polymerase...several micrograms of target DNA from just a few nanograms of template DNA through several cycles of denaturation...this step enables the DNA polymerase to synthesize and elongate the new target DNA strand accurately and... are using the same template DNA for all your reactions, the Template DNA can be added to the master mix...the number of nanomoles of DNA times 10. For example, if your lyophilized DNA is 38.5nm, add 385µl of water...

  8. Lentivirus Production

    Type
    Protocol
    ...Transfection grade DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and Transfer plasmid encoding your...therefore, plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, ...transfection plasmids into OptiPro SFM: Reagent Amount per 10 cm dish* Packaging Plasmid ( psPAX2 ) 1.3...1.3 pmol Envelope Plasmid ( pMD2.G ) 0.72 pmol Transfer Plasmid* 1.64 pmol OptiPro SFM to total volume...volume 500 μL *Plasmid concentrations and ratios should be optimized for each transfer plasmid. Pro-Tip Endotoxins...dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would be 83.4 μL of 1 mg/mL PEI in 416.6 μL of OptiPro...the diluted DNA mixture. Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate...

  10. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...taken up the plasmid. This can serve as an alternative to antibiotic selection. Plasmids of this type .... Scientists can easily utilize GFP-containing plasmids as a means to many functional ends. We’ve listed...each protein or domain being tested. Find FRET plasmids here. Split EGFP: An alternative to FRET, split... marking/selection: Expression constructs like plasmids often include GFP as a marker to help identify...nude mice in cancer studies, act as a reporter for DNA double strand break repair, and to label pathogenic...techniques in fluorescent microscopy. Read More: All Plasmids 101 Articles Addgene's Fluorescent Protein Guide... plasmids 101...

  11. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...sequence in pEcgRNA. Next both plasmids are transformed in E. coli with the donor DNA for editing to take place...beneficial for plasmid clearance. It is more convenient to update the spacer(20nt) on pEcgRNA plasmid as only.... pCutamp cures the plasmids by targeting the ampR promoter in all three plasmids. pCutamp is then removed... 13 plasmids with 14 rounds of transformation in 30 days. Figure 2: (A) MUCICAT plasmids for ... original pCas/pTargetF system consists of one plasmid that expresses the S. pyogenes Cas9, a temperature-sensitive...the pTargetF construct and then introduce both plasmids into the cell. The edit in the chromosome can ...the pMB1 replicon, resulting in cutting of the plasmid. At this point, subsequent introductions of pTargetF...

  12. Celebrating 15 Years of Scientific Sharing

    Type
    Blog Post
    ...the challenges of DNA distribution to ensure scientists everywhere can access plasmids despite the evolving... that is something to recognize! Distributing plasmids, viral service, and more For the first time ...to worry about succeeding at our core mission. Plasmid distribution is amazingly still growing at >9% ...Addgene Resources on the Addgene website Find plasmids for your research  Learn about our viral service...

  13. Protocol - How to Streak a Plate

    Type
    Protocol
    ...for DNA purification will minimize the chance of having a mixture of plasmids in your purified DNA. This...Agar Plates Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture Introduction If you have... stab culture of bacteria and need to purify plasmid DNA from it, you will want to isolate an individual...single colonies, you can proceed to Recovering Plasmid DNA or use the individual colonies for other experiments...antibiotic. Label the bottom of the plate with the plasmid name and the date. It is also a good idea to add...overnight (12-18 hours) at 37 °C. Pro-Tip Some plasmids or bacteria need to be grown at 30 °C instead ... 37 °C. This is often true for large unstable plasmids, which sometimes recombine at 37 °C. Be sure to...

  14. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    ... How to Use the Nanobody Toolbox Find Nanobody Plasmids Structural comparison As the name suggests, ... Species Antibody Subclass Nanobody Addgene Plasmid Rabbit N/A* TP897 pTP1183 (1x cysteine) pTP955...Nanobody In Bacteria: Table 2 also has a link to the plasmid you’ll need to express your nanobody of choice ...column. bdNEDP1 protease can be generated using plasmid pDG02583. Label the Nanobodies: Purified nanobodies...Journal of Cell Biology. PubMed PMID: 29263082 Find plasmids from this publication at Addgene. 2. Bates, M...23495938. Additional Resources on the Addgene Blog DIY DNA ladders from Penn State University Use Nanobodies... on Addgene.org Find more nanobody expression plasmids here Additional Resources Looking for monoclonal...

  15. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...episomal expression. Episomes are circular pieces of DNA that reside outside a host cell’s genome. In this... The most obvious workaround is to use a multi-plasmid system, where you have one AAV vector dedicated...levels of AAV vector integration into CRISPR-induced DNA breaks. Nature Communications, 10(1). https://doi.org...

  16. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...derivatives. Unmodified Cas9 causes double stranded DNA breaks that commonly result in a complete loss of...These mutations result in an inactive RNA-guided DNA binding protein (dCas9). dCas9 is then tethered to... CRISPR Resources on Addgene.org Find CRISPR plasmids and resources here Find pooled libraries available...viability genes as they are subject to an increase in dsDNA breaks that can cause cell death. Thus, CRISPR inhibition...mitigating toxicity induced by high copy number dsDNA cutting. A Dolcetto library containing only 3 sgRNAs...

  17. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...random) integration into the genome, and its large DNA packaging capacity is well suited for larger transgenes...AAV. Its main limitation is the small size of its DNA cargo - 4.8 kb at most - which means that a gene ...in mammals Resources at Addgene.org Find AAV plasmids Find ready-to-use AAV viral preps Browse all viral...

  18. The Challenges of Cell Culture

    Type
    Blog Post
    ...good that it is used as a forensic technique (aka DNA fingerprinting) that can place suspects at the scene...range of testing, from animal doping to forensics, DNA sequencing to drug testing. You can find Nick on ...Protein Localization Resources on Addgene.org Find Plasmids for Mammalian Cells ...

  19. Video Library

    Type
    Protocol
    ...Storing and Handling Addgene Plasmids What to do after you receive your plasmids from Addgene Instructions...Protocol Purifying DNA from an Agarose Gel Gel purification allows you to isolate and purify DNA fragments based...electrophoresis, you can cut DNA bands out of the agarose gel and purify the DNA samples. This is a commonly...Protocol Diagnostic Restriction Digest Verify a plasmid using restriction enzymes Diagnostic Restriction...Instructions for Storing and Handling Plasmids Genomic Deletions with CRISPR Generate genomic deletions in mammalian...Transformations We walk you through a standard plasmid transformation protocol and offer some tips and...the standard laboratory procedure for separating DNA by size for visualization and purification. Agarose...

  20. FlipGFP, a novel fluorescence protease reporter to study apoptosis

    Type
    Blog Post
    ... external signals such as developmental cues and DNA damage. These signals activate a cascade of caspases...these diseases and disorders. You can find the plasmids for FlipGFP and FlipCherry at Addgene. References...Download the fluorescent proteins 101 eBook Read our plasmids 101 blog series Resources on Addgene.org Browse...protein fusions Browse the fluorescent proteins plasmid kits ...
Showing: 321 - 340 of 509 results