Skip to main content
Addgene

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

Information

In Category

We narrowed to 36 results for: Rest

Showing: 1 - 10 of 36 results

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

    Type
    Protocol
    ...Plasmid Cloning by Restriction Enzyme Digest (aka Subcloning) You may also like... Restriction Digest of Plasmid...identify which restriction sites are present in a given sequence. When selecting restriction enzymes, you...desired restriction sites to flank your insert : You can use PCR Based Cloning and add restriction sites...need to avoid restriction enzymes that cut within your insert. Adding desired restriction sites to your...Information about plasmid cloning by restriction enzyme digest (subcloning), including design and experimental... another as long as it is already bounded by restriction sites that are also present in the same orientation...Backbone Reference . Background Subcloning by restriction digest is a commonly used lab technique. For ...

  2. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ... Protocols Restriction Digest of Plasmid DNA Restriction Digest...commonly used restriction enzymes, visit (Link opens in a new window) NEB . Restriction enzyme digestion...of your plasmid of interest (see below for recommend amounts) Appropriate restriction enzyme (see manufacturer's...Procedure Select restriction enzymes to digest your plasmid. Pro-Tip To determine which restriction enzymes will...typical restriction digestion reaction could look like this: 1 µg DNA 1 µL of each Restriction Enzyme ...perform a restriction digest of plasmid DNA Protocols...from an Agarose Gel DNA Ligation Introduction Restriction enzyme digestion takes advantage of naturally...

  3. Kit Free RNA Extraction

    Type
    Protocol
    ...minutes at 12,000 × g at 4°C to separate RNA from the rest of the tissue/cell lysate. Pro-Tip Having your samples...

  4. Protocol - Bacterial Transformation

    Type
    Protocol
    ...recommend that you plate 50 μL on one plate and the rest on a second plate. This gives the best chance of...also like... Plasmid Cloning by Restriction Enzyme Digest Restriction Digest of Plasmid DNA DNA Ligation... how to transform E. coli with your plasmid of interest. Protocols...

  5. Pipetting Protocol

    Type
    Protocol
    ...release the plunger, allowing it to go back to its resting position. Wait for the liquid to stop flowing into...

  6. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ... or by RT-PCR. Isolate your PCR product from the rest of the PCR reaction using a kit, such as the (Link...assist with restriction enzyme digestion (usually 3-6bp). Restriction Site: Your chosen restriction site for...fusing the upstream restriction site to the forward primer and the downstream restriction site to the reverse...product is now ready for restriction digestion. Digest Your DNA Set up restriction digests for your PCR product... Plasmid Cloning by PCR You may also like... Restriction Digest of Plasmid DNA DNA Ligation Bacterial ...of a piece of DNA and at the same time adding restriction sites to the ends of that piece of DNA so that...that it can be easily cloned into a plasmid of interest. For this example, we will describe how to copy...

  7. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...Diagnostic Restriction Digest Diagnostic Restriction Digest You may also like... Restriction Digest of ...another. A diagnostic restriction enzyme digest takes advantage of the fact that restriction enzymes cleave ... your plasmid of interest (see below for recommended amounts) Appropriate restriction enzyme (see manufacturer's...Orientation by Restriction Digest If you clone an insert into a vector by a single restriction enzyme, you...DNA Agarose Gel Electrophoresis Introduction Restriction enzymes are naturally occurring bacterial endonucleases...enzymes and the unique sites they recognize, restriction digests have become the most widely used method...cleave DNA at specific sequences called restrictions sites. Often, the size of the plasmid insert and vector...

  8. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ... spectrophotometer Restriction Digests Cut and analyze plasmid DNA with restriction enzymes Agarose Gel...pooled-plasmid libraries Diagnostic Restriction Digest Verify a plasmid using restriction enzymes Watch the Video!...new window) Link to Video Restriction Cloning Assemble plasmids by restriction digest and ligation Cloning...Cloning by PCR Generate restriction sites by PCR Modification by Annealed Oligo Cloning Add a short stretch... Cloning (LIC) Scarless cloning with Type II restriction enzymes and T4 polymerase pLKO.1 - TRC Cloning...

  9. Western Blot

    Type
    Protocol
    ...protein of interest, and a negative control sample that does not express the protein of interest....detection using antibodies specific to the protein of interest. This protocol describes the basic steps for lysing... Precast SDS-PAGE gel SDS-PAGE running buffer Prestained protein ladder Ethanol iBlot 2 PVDF Mini Stack...depending on the cellular location of the protein of interest. RIPA buffer is suitable for most proteins but...the samples into the gel. Load 5–10 µL of the prestained protein ladder. Place the lid on the tank and...sample type and cellular location of the protein of interest. You may need to try a variety of lysis buffers...

  10. Plasmid Modification by Annealed Oligo Cloning (with Protocols)

    Type
    Protocol
    ... Annealed Oligo Cloning You may also like... Restriction Digest of Plasmid DNA DNA Ligation Bacterial ...this tutorial we will discuss how to add new restriction sites to the MCS of an empty vector. However,...design a set of oligos containing your desired restriction sites and add them to your existing vector. It...cloned directly into the overhangs generated by restriction digest of existing sites in the original vector...waiting for your oligos to arrive, conduct a restriction digest of 1μg of vector with EcoRI and SalI. ...
Showing: 1 - 10 of 36 results