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  1. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...sequence for degradation. Viral introduction of shRNA plasmids can induce gene-specific silencing temporarily...transfection and random genomic integration of a plasmid. Viral transduction is also generally less toxic...
  2. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    ...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...
  3. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...recipient plasmid (usually in the Multiple Cloning Site (MCS)), but do not cut elsewhere on the plasmid Will...sites to your recipient plasmid : You can modify the MCS of your recipient plasmid using Annealed-oligo ...material. We recommend 1.5-2μg of donor plasmid and 1μg of recipient plasmid. It is also critical that as much...re-circularization of the recipient plasmid. You should treat your digested recipient plasmid with a phosphatase prior...ligations for each plasmid you are trying to create, with varying ratios of recipient plasmid to insert. It...recipient plasmid + insert plate should have significantly more colonies than the recipient plasmid alone ... on your recipient plasmid alone plate, you can try ligating the recipient plasmid alone in the presence...
  4. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...for verifying plasmids like this is "plasmid fingerprinting", where you cut the plasmid into 3-8 pieces...used to verify your plasmid. The goal of a diagnostic digest is to cut your plasmid into specific sized...insert and the 5000bp backbone. Verifying Plasmid Identity by "Plasmid Fingerprinting" Often, it will be enough...for how to perform a diagnostic digest to verify plasmid size, verify insert orientation, and more. Protocols...Digest You may also like... Restriction Digest of Plasmid DNA Agarose Gel Electrophoresis Introduction Restriction...selectively move a specific piece of DNA from one plasmid to another. A diagnostic restriction enzyme digest...called restrictions sites. Often, the size of the plasmid insert and vector backbone are known and thus this...
  5. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ... Protocols Plasmid Cloning by PCR Plasmid Cloning by PCR You may...recipient plasmid (usually in the Multiple Cloning Site (MCS)), but do not cut elsewhere on the plasmid. Bonus...recipient plasmid. Remember to insert your DNA in the correct orientation in the recipient plasmid by viewing... 1μg of recipient plasmid. It is also critical that as much of the recipient plasmid as possible be cut...ligations for each plasmid you are trying to create, with varying ratios of recipient plasmid to insert. It...recipient plasmid + insert plate should have significantly more colonies than the recipient plasmid alone ... on your recipient plasmid alone plate, you can try ligating the recipient plasmid alone in the presence...
  6. Protocol - How to Inoculate a Bacterial Culture

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    Protocol
    ...Storage of Plasmids Recovering Plasmid DNA from Bacterial Culture Background Information Plasmids can carry...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... isolate your plasmid DNA from the bacterial culture by following Isolating Your Plasmid DNA . Antibiotic... 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 ...clonal) colonies of bacteria carrying a specific plasmid. However, a liquid culture is capable of supporting...
  7. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    ...photoswitchable kinases in my experiments? You can find the plasmids from Zhou et al here. In addition, it could be...GFP Resources at Addgene.org Find KTR reporter plasmids Find photoactivatable fluorescent proteins in ...
  8. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ...Resources Additional resources on the Addgene blog Plasmids 101: Aptamer FluorophoresAntibodies 101: scFVs...available from AddgeneBrowse our recombinant antibody plasmid collection References Grumezescu, A. M. (2018)....
  9. Choosing the B(right)est Fluorescent Protein: Photostability

    Type
    Blog Post
    ...localized fusion proteins and do not require dedicated plasmids or constructs. It should be noted that the photostability...Find Empty Backbones with Fluorescent Tags Find Plasmids for Subcellular Localization Browse the entire...
  10. New Videos: Addgene Lab Tips

    Type
    Blog Post
    ...Addgene has distributed over 800,000 plasmids since 2004. In that time, our lab has gotten a lot experience...
  11. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...Recovering Plasmid DNA from Bacterial Culture Recovering Plasmid DNA from Bacterial Culture You may also...concentrated plasmid DNA. This page will discuss the general procedure for purifying plasmid DNA from bacterial...perform plasmid purification without using a kit, you can find a protocol for kit-free plasmid mini-prep... the plasmid-containing supernatant. Add either ethanol or isopropanol to precipitate the plasmid DNA....high concentration of clean plasmid DNA. To improve the purity of plasmid DNA purified without a kit it...How to purify plasmid DNA from a bacterial culture, including protocols, tips, and FAQ Protocols... a new window) Promega sell kits for isolating plasmid DNA in quantities as low as a few micrograms to...
  12. Protocol - Bacterial Transformation

    Type
    Protocol
    ... also like... Plasmid Cloning by Restriction Enzyme Digest Restriction Digest of Plasmid DNA DNA Ligation...both storing and replicating plasmids. Because of this, nearly all plasmids (even those designed for mammalian...that will help to maintain the plasmid without rearrangement of the plasmid DNA. Additionally, specific ...tube of plasmid DNA and just need to transform bacteria so that you can grow up more of the plasmid) you ...efficient at taking up larger plasmids. If you need to transform large plasmids, it is a good idea to use ...Learn how to transform E. coli with your plasmid of interest. Protocols...introduced into a cell. Transformation of bacteria with plasmids is important not only for studies in bacteria ...
  13. 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... Storage of Plasmids Creating Bacterial Glycerol Stocks for Long-term Storage of Plasmids You may also...important for long-term storage of plasmids. Although you can store your plasmid DNA at -20°C, many labs also...also like... Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture Background Information... and growth temperature, can be found on your plasmid's information page. The next day you will be able...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...
Showing: 481 - 500 of 883 results