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Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

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We narrowed to 26 results for: SIM

Showing: 1 - 20 of 26 results

  1. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...ImageJ or similar software Reagents Purified recombinant antibody, 0.9–1.1 mg/mL SimplyBlue SafeStain,... bio-safe Coomassie G-250 stain (Addgene uses SimplyBlue SafeStain) and ImageJ software. The sample is...and reagents from other vendors should produce similar outcomes when using these protocols. However, please... Pour off the water in the sink. Add 20 mL of SimplyBlue SafeStain and incubate for 1 h with gentle agitation...agitation on a rocking platform. Pour off the SimplyBlue SafeStain in the sink. Add 100 mL of deionized...Analysis Use ImageJ (Link opens in a new window) or a similar photo software to determine the relative intensity...

  2. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...cloning sites, will result in similar sized bands, thus making this simple digest less informative. This...Plasmid Size -OR- Insert and Backbone Size The simplest form of diagnostic digest is one in which you ...products from each orientation are different sizes. Simply run the digest products on a gel and those clones...

  3. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...choice with minimal limitations. Background In its simplest form, PCR based cloning is about making a copy...TAAGCAGAATTCATGTGGCATATCTCGAAGTAC-3'. For the Reverse Primer, the design is similar, but we need to use the reverse complement to get... get PCR amplification. We can start similarly, taking the final 18bases of the ORF, including the stop...primer. If you are amplifying from a plasmid or simple template, there is very little chance for mis-priming...

  4. Kit Free RNA Extraction

    Type
    Protocol
    ... (Sarkosyl) 0.1 M 2-mercaptoethanol TRIzol® or similar product such as TRI Reagent®, RNAzol®, QIAzol® ...how effective your cell lysis protocol is. While simple homogenization is effective for most mammalian ...Homogenize or lyse tissues or cells in TRIzol® or a similar product. For tissues: use 1 mL of TRIzol® per 100...

  5. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...bands needed to be separated - see FAQs below . Simply adjust the mass of agarose in a given volume to...How do you get better resolution of bands? A few simple ways to increase the resolution (crispness) of ...you get better separation of bands? If you have similarly sized bands that are running too close together...

  6. Protocol - How to Design Primers

    Type
    Protocol
    ...primers are mainly used for amplifying a small, simple fragment of DNA. On the other hand, a long primer...The structure of the primer should be relatively simple and contain no internal secondary structure to ...

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

    Type
    Protocol
    ...do you get better resolution of bands? A couple simple ways to increase the resolution (crispness) of ...you get better separation of bands? If you have similarly sized bands that are running too close together...

  8. Ligation Independent Cloning

    Type
    Protocol
    ... Your Primers Primer design for LIC is often as simple as using the backbone manufacturer's suggested ...the first G on 3'-5' strand from cut site). For simplicity, only the 5' primer is shown here. Note: Use ...

  9. Centrifugation

    Type
    Protocol
    ...RNA extractions. Plus, centrifuges are useful for simply collecting liquid to the bottom of a tube. You ... can be about bench height. They often spin at similar speeds to their smaller cousins, but can hold much...

  10. Gibson Assembly Protocol

    Type
    Protocol
    ... al. , 2010). Number of fragments assembled simultaneously Multiple fragments can be assembled in one ...opens in a new window) Rabe BA, Cepko C. (2020). A Simple Enhancement for Gibson Isothermal Assembly. bioRxiv...

  11. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...can create thousands of identical copies using a simple set of reagents and a basic heating and cooling...Your 5’ and 3’ primers should be designed to have similar melting temperatures (Tm). Set the annealing temperature...

  12. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...and reagents from other vendors should produce similar outcomes when using these protocols. However, please...plate washer (optional) Scale Microsoft Excel or similar software Reagents Purified antigen Purified recombinant...

  16. Video Library

    Type
    Protocol
    ...pipette and avoid inconsistent pipetting with our simple tips Bacterial Transformations We walk you through...
Showing: 1 - 20 of 26 results