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

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Showing: 1 - 20 of 23 results

  1. Gibson Assembly Protocol

    Type
    Protocol
    ...in an assembly if you want to save time. Working on ice, combine segments in the Gibson Assembly Reaction... Number of fragments assembled simultaneously Multiple fragments can be assembled in one reaction. However...Gibson Assembly (Link opens in a new window) OpenWetWare: Janet Matsen’s guide to Gibson Assembly (Link...Get started with Gibson Assembly Cloning! Protocols... Protocols Gibson Cloning Gibson Assembly Cloning You may also ...Institute developed a novel method for the easy assembly of multiple linear DNA fragments (Gibson et al...three different enzymes, the product of a Gibson Assembly is a fully ligated double-stranded DNA molecule...

  2. Western Blot

    Type
    Protocol
    ...Block the membrane in blocking buffer for 1 h at RT on a shaking platform. Wash the membrane 3x for 5 ...Incubate the membrane with secondary antibody for 60 min at RT on a shaking platform. Wash the membrane 3x for...incubate the membrane in the chemiluminescence reagent for 5 min at RT . Cover the membrane with clear ...ethanol into 8 mL of deionized water and mix well. Assemble the transfer sandwich as follows: Unseal the iBlot...in deionized water and place it on the transfer membrane of the bottom stack. Soak a piece of iBlot Filter...information before running your experiment. Remove the membrane from the transfer device and place it in a container... is typically between 1–10 μg/mL. Incubate the membrane overnight in primary antibody at 4 °C on a rocking...

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

    Type
    Protocol
    ...on an agarose gel Watch the Video! DNA Ligation Assemble DNA using DNA ligase Bacterial Transformation ... new window) Link to Video Restriction Cloning Assemble plasmids by restriction digest and ligation Cloning...Add a short stretch of DNA to a plasmid Gibson Assembly Combine overlapping DNA fragments in a single ...

  4. AAV Production in HEK293 Cells

    Type
    Protocol
    ...drifted. Filter the solution through a 0.22 μm membrane. Aliquot 500–1000 μL into sterile tubes. Store... pH to ~8.5 Filter sterilize through a 0.22 μm membrane. Store at 4 °C. Considerations Before You Start...supernatant as follows: Filter through a 0.45 μm PES membrane. Add 40% PEG such that the final PEG concentration...

  5. General Transfection

    Type
    Protocol
    ...drifted. Filter the solution through a 0.22 µm membrane. Aliquot 500–1000 µL into sterile tubes. Store...limited effect on cell growth. Last reviewed on: November 7, 2023...

  6. Protocol - Bacterial Transformation

    Type
    Protocol
    ... still get enough colonies for your next step. Remember that each of these shortcuts will reduce the efficiency...field is applied to the cell/DNA mixture to induce membrane permeability. To do this you will need to have...

  7. Using a Light Microscope Protocol

    Type
    Protocol
    ...started in the lab! Introduction Microscopes are emblematic of biological research and are found in many ...focus. Pro-Tip As you increase your magnification, remember that the objectives are larger. Adjust your focus...

  8. Pipetting Protocol

    Type
    Protocol
    ...pipette tip and using the pipette. Last Update: September 2022 Video Watch the video for tips on pipetting...preloaded into tip boxes. In either case, once assembled into their boxes, pipette tips can be sterilized...

  10. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...Vector Addgene Plasmid 10878. Protocol Version 1.0. December 2006. Copyright Addgene 2006, All Rights Reserved... . 1996 272:263-267. PubMed . Asymmetry in the assembly of the RNAi enzyme complex. . Schwarz DS et. al...

  12. Water Bath Protocol

    Type
    Protocol
    ... a water bath with individual slots for tubes, remember that when you add the tube to the slots, it can...

  13. Ligation Independent Cloning

    Type
    Protocol
    ...following T, so that there are no gaps in the final assembly. In this case, 15 bp of homologous sequence is...

  14. Centrifugation

    Type
    Protocol
    ...your lab’s instrument before use. Last Update: September 2022 Equipment Centrifuge Microfuge tubes or other...

  17. Lentivirus Production

    Type
    Protocol
    ...drifted. Filter the solution through a 0.22 μm membrane. Aliquot 500–1000 μL into sterile tubes. Store...

  20. Pouring LB Agar Plates

    Type
    Protocol
    ...with your antibiotic of interest. Last Upload: September 14, 2016 Protocol Video Before you begin this ...
Showing: 1 - 20 of 23 results