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Addgene

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

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We narrowed to 47 results for: TIM

Showing: 21 - 40 of 47 results

  1. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ...double digest (digesting with two enzymes at the same time), you will need to determine the best buffer that...and the amount of DNA in the reaction, incubation time can range from 45 mins to overnight. For diagnostic... of methylation sensitive restriction sites . Sometimes enzymes cut sequences which are similar, but not...amount of Master Mix to each tube. This will save you time and ensure consistency across the reactions....

  2. Colony Formation Titering Assay

    Type
    Protocol
    ...the same time as the virus-mediated transduction. For some cell lines, transduction is optimized if the ...cannot be used for later experiments. Workflow Timeline Day 0: Seed and transduce cells Day 2: Replace...that case, polybrene should only be added at the time of viral transduction, and not during the cell seeding...

  3. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...column gradient for AAV purification. Workflow Timeline Day 1: Purify Day 2: Buffer exchange and concentration... a T70i rotor at 10 °C. Pro-Tip If you need more time, you can alternatively centrifuge for 2 h at 200,000...Repeat for each QuickSeal tube. (Optional) For first time users, it is a good idea to assay each fraction ...sample and make sure to pipet back and forth a few times to mix the iodixanol that has settled at the bottom...

  4. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ...through the use of a selection curve to determine optimal antibiotic concentration. Last Update: Oct. 27,...30 minutes with the lid on to give the antibiotic time to more fully absorb. During the incubation, transform...Shown below are the results from an experiment optimizing the concentration of carbenicillin, plated over-agar...

  5. Water Bath Protocol

    Type
    Protocol
    ...liter. Place a thermometer in the water bath. Oftentimes, the water bath will have a spot to secure the...that it will reach the desired temperature by the time you need it for your experiment. Water baths tend... the water bath, place the lid back on. Note the time that you’ve put your materials in the water bath...

  6. Lab Safety for Biosafety Levels One and Two

    Type
    Protocol
    ...protective equipment (PPE) and wear it the whole time you are working in the lab. Do not eat, drink, chew...supervisor what is available and the appropriate contact times for each agent. Make sure you have enough space ...laboratory. Don’t mouth pipette! There may be some times when you may be working with a protocol that requires...

  7. Using a Light Microscope Protocol

    Type
    Protocol
    ...magnifies the image four times the actual size, the 10x objective magnifies it 10 times, and so on. The ocular...focus, use the stage arm to move the stage (and ultimately your slide) until your field of view is centered...

  8. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...approximately 75-80% of the way down the gel. A typical run time is about 1-1.5 hours, depending on the gel concentration... use long-wavelength UV and expose for as little time as possible to minimize damage to the DNA. Note:...the gel at a lower voltage for a longer period of time; b) using a wider/thinner gel comb; or c) loading...

  9. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ... of Solution II and invert the tube carefully 5 times to mix the contents. The contents will become clear...Solution III to each tube. Mix by inverting several times. A white precipitate will be formed which contains... 0.1 mM EDTA). Pro-Tip DNA resuspension can take time, it is a good idea to let it sit for several hours...

  11. Protocol - How to Streak a Plate

    Type
    Protocol
    ...it through a flame, just be sure to allow enough time for the loop to cool before touching it to the bacteria...often true for large unstable plasmids, which sometimes recombine at 37 °C. Be sure to check this before...

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

    Type
    Protocol
    ...will want to use long-wavelength UV for as short a time as possible to get the bands cut out. Once you have...the gel at a lower voltage for a longer period of time; b) using a wider gel comb; or c) loading less DNA...

  13. Weighing Reagents Protocol

    Type
    Protocol
    ...weigh out 100 g three times, placing the sucrose in your container after each time. Place the weighing ...

  14. Ligation Independent Cloning

    Type
    Protocol
    ...Independent Cloning (LIC) obviates the need for the time-consuming ligation step of traditional cloning methods...sterile water (instead of TE buffer) to ensure optimal salt concentrations in subsequent reactions. Step...

  15. Centrifugation

    Type
    Protocol
    ...place, and the control panel where you can set the time and speed needed for your experiment. Procedure ...Adjust the centrifuge settings according to speed and time (and temperature, if applicable) listed in your ...

  16. Fluorescence Titering Assay

    Type
    Protocol
    ... institution’s biosafety regulations. Workflow Timeline Day 0: Seed 293T cells Day 1: Transduce cells ...with multiple integration events leading to underestimation of the true titer. Calculate the transduction...

  17. Gibson Assembly Protocol

    Type
    Protocol
    ...can work fine in an assembly if you want to save time. Working on ice, combine segments in the Gibson ...rate when assembling more than five fragments at a time. Resources and References New England Biolabs (NEB...

  18. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ... making a copy of a piece of DNA and at the same time adding restriction sites to the ends of that piece...both function in the same buffer, as this will save time later. In our example, we will use EcoRI and NotI...

  19. DNA Quantification

    Type
    Protocol
    ... the absorbance of a liquid you can accurately estimate the concentration of a substance in that liquid...
Showing: 21 - 40 of 47 results