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

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We narrowed to 987 results for: des

Showing: 921 - 940 of 987 results

  2. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...restriction enzyme digest (subcloning), including design and experimental procedures....easily move YGOI into a mammalian expression vector. Design (Choosing enzymes) Many DNA analysis tools, including..., but do not cut within your insert Are in the desired location in your recipient plasmid (usually in ...your recipient plasmid as well as a specifically designed test digest later to verify that the insert was... fear. You have other options, such as: Adding desired restriction sites to flank your insert : You can...restriction enzymes that cut within your insert. Adding desired restriction sites to your recipient plasmid : You... orientation of your insert, so you may want to design a diagnostic digest for this purpose. Congratulations...

  3. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...Vector Guides Virus Blog Posts Addgene Protocols Viral Service Introduction This protocol describes how ...This protocol describes how to generate a monoclonal cell line from a polyclonal pool of stable cells...type will be able to grow under the conditions described in this protocol. Conditioned medium (details ... specific conditions used. Each 96-well plate described in this protocol could, in theory, give rise to...protocol was performed at Addgene on the A549 cells described in our sample data , each 96-well plate gave rise...from Feng Zhang (Addgene plasmid #52962 ) and is described in Improved vectors and genome-wide libraries ...

  8. Ligation Independent Cloning

    Type
    Protocol
    ...experimental design. Search Addgene's collection of empty LIC cloning vectors Protocol Step 1: Design Your Primers... Primers Primer design for LIC is often as simple as using the backbone manufacturer's suggested leader...primer is shown here. Note: Use web-based primer design software to ensure a melting temperature between...nucleotide dGTP in the reaction (exclude all other nucleotides from standard polymerase protocol), causing the...manufacturer. It is very important to remove all free nucleotides from your PCR product before proceeding, as they...

  11. How to Be an Excellent Trainee

    Type
    Blog Post
    ...will identify a ‘scientific mentor’: someone who provides career advice and offers direction on your scientific...

  14. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...plasmid of interest. For this example, we will describe how to copy a cDNA from one vector into a new ...(YGOI) for ligation into a recipient plasmid. Designing Primers for PCR Based Cloning The basic PCR primers...that: Do not cut within your insert. Are in the desired location in your recipient plasmid (usually in ...examine the DNA sequence that we want to amplify and design primers that will bind to and replicate it. The...ends of the ORF and how these are used for primer design: Because we are cloning an ORF, we want to clone...TAAGCAGAATTCATGTGGCATATCTCGAAGTAC-3'. For the Reverse Primer, the design is similar, but we need to use the reverse complement...

  15. Western Blot

    Type
    Protocol
    ...This protocol describes the basic steps for lysing cells, determining total protein concentration in ...specific to the protein of interest. This protocol describes the basic steps for lysing cells, determining ...continue running the gel until you have obtained the desired separation. Gently open the gel with the spatula...Device. Close the lid of the device. Select the desired method and make sure the parameters are correct... incubation Dilute the primary antibody to the desired concentration in blocking buffer. Pro-Tip The optimal...peroxidase-conjugated secondary antibody to the desired concentration in blocking buffer. Pro-Tip The optimal...

  16. Protocol - How to Perform Sequence Analysis

    Type
    Protocol
    ...primer should be a minimum of 50 nucleotides and a maximum of 300 nucleotides from your target. Are there ...Analysis of a Plasmid You may also like... How to Design a Primer Diagnostic Restriction Digest Introduction...If you require additional sequencing and need to design a custom primer, Addgene recommends using Addgene's...'s sequencing results as a reference for primer design. A good sequencing reaction will produce between...

  17. Water Bath Protocol

    Type
    Protocol
    ... of bacteria or fungi. There are disinfectants designed to be used in water baths with instructions on...bath on and set the appropriate temperature as described in your protocol. How to set the temperature will...Set up your water bath so that it will reach the desired temperature by the time you need it for your experiment...water bath to prevent evaporation and maintain the desired temperature. This also helps the water bath heat...water bath even if you are using disinfectants as described above in step 3. You will also need to maintain...

  18. Five Popular Model Organisms

    Type
    Blog Post
    ...difficult and time consuming. GAL4/UAS was first described in 1993 by Norbert Perrimon’s lab and has been...
Showing: 921 - 940 of 987 results