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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 1,086 results for: ANT;

Showing: 1061 - 1080 of 1086 results

  1. Water Bath Protocol

    Type
    Protocol
    ... tub. Disinfectant may be added to prevent growth of bacteria or fungi. There are disinfectants designed...Bath 10% bleach or 70% ethanol Distilled water Disinfectant Thermometer Water bath weights and floats Reagents...allow the temperature to stabilize. Use water resistant markers, such as permanent markers, to label your...water bath, be sure to use a marker that is also resistant to chemicals such as ethanol. Writing on the tops...clean out the water bath even if you are using disinfectants as described above in step 3. You will also ...

  2. Protocol - How to Streak a Plate

    Type
    Protocol
    ...appropriate antibiotic) Bacterial stab Procedure Obtain an LB agar plate with appropriate antibiotic. Label...add the antibiotic resistance and your initials. Labeling within a laboratory setting is important for organization...and need to purify plasmid DNA from it, you will want to isolate an individual clonal population (single...

  3. Centrifugation

    Type
    Protocol
    ... save the pellet, the supernatant, or both. Conclusion Centrifuges are important tools for many different...volume as your samples. If your samples are significantly higher density than water, then you may need... The centrifuge should come up to speed with a pleasant hum. If you hear loud noises or the centrifuge...sample while the liquid portion is known as the supernatant. Depending on your experiment, you may be required...

  4. AAV Production in HEK293 Cells

    Type
    Protocol
    ...on ice and transfer supernatant to a sterile 500 mL bottle. Process the supernatant as follows: Filter ... Add 25 mL of PEG solution to each 100 mL of supernatant. Split into 2 x 500 mL sterile bottles as needed... at 3900 rpm for 15 min at 4 °C. Discard the supernatant and resuspend the pellets in a total of 5 mL ...now ready to purify your prep . The clarified supernatant can be kept overnight at 4 °C before proceeding...

  5. Protocol - How to Create a Bacterial Glycerol Stock

    Type
    Protocol
    ...Background Information Bacterial glycerol stocks are important for long-term storage of plasmids. Although you...glycerol stocks of their plasmids. This way, when you want to make more plasmid DNA, the plasmid will already...be stored at 4°C for a few weeks. However, if you want to store bacteria for a longer time, you will need...completely and will improve the shelf life. It is very important that you shake the glycerol before freezing (5...

  6. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ... A diagnostic restriction enzyme digest takes advantage of the fact that restriction enzymes cleave DNA...form of diagnostic digest is one in which you just want to verify that the plasmid that you have is the ...the correct orientation. In the example below we want to know how to differentiate between two clones...or enzymes) that cuts within the insert, but significantly off center (ideally around 1/3 of the way from...

  7. Using a Light Microscope Protocol

    Type
    Protocol
    ... As the name suggests, light microscopes take advantage of the physical properties of light to detect ...detect small objects. Two of the most important properties of a microscope are magnification (the ability to...see several pieces of information, but the most important (for now) is the magnification power, such as ...your images in the future. Conclusion Like any important laboratory instrument, you should be sure to take...

  8. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...been sufficiently selected with the appropriate antibiotic so that every cell in the culture has, in theory...maintain cells in selective medium at a reduced antibiotic concentration (e.g., half the concentration that...Sample Calculation First, use a hemocytometer to quantitate the homogenized cell solution. Homogenized cell... additional 3 weeks under blasticidin selection. Anti-Cas9 Western blotting was performed on whole-cell...

  9. Lentivirus ddPCR Titration

    Type
    Protocol
    ...Centrifuge for 5 min at 100 x g . Gently aspirate supernatant. Wash cell pellets in 500 µL PBS. Centrifuge ...Centrifuge for 5 min at 100 x g . Gently aspirate supernatant. Extract genomic DNA according to the GeneJet Genomic...transfer the plate to the Droplet Reader. Open the QuantaSoft software to set up a new plate layout. Designate...the dilutions. RPP30 copies should be relatively constant across samples. In the RRE example below, 2-fold...

  10. Personal Protective Equipment (PPE) for BSL-1 and BSL-2 Labs

    Type
    Protocol
    ... in healthy humans. Many of the precautions are meant to protect individuals from potential accidents ...don’t touch your face! Close-toed shoes and long pants should be worn to protect your legs and feet from...the safest way possible. Protecting yourself is important! Disclaimer: The SOP presented here has been designed...

  11. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ...Introduction Restriction enzyme digestion takes advantage of naturally occurring enzymes that cleave DNA...given digestion will depend on the amount of DNA you want to cut. By definition: one unit of enzyme will cut...Dam or Dcm methylase positive strains will be resistant to cleavage at certain restriction sites. See ...

  12. Lentivirus Production

    Type
    Protocol
    ...harvesting. Filter supernatant through a 0.45 μm PES filter. The viral supernatant can be stored at 4 ...at 4 °C between harvest. Centrifuge the viral supernatant at 2100 rcf for 5 minutes to pellet any packaging...

  13. Pipetting Protocol

    Type
    Protocol
    ...measurement in the next step. The second stop is meant to expel excess liquid from the tip once you’ve ...Conclusion Pipetting accurately and precisely is important since even small changes in the volume could affect...Beyond knowing the proper way to pipette, it’s important for scientists to calibrate their pipettes to ...

  14. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...Note: If you add EtBr to your gel, you will also want to add it to the running buffer when you run the...separate larger bands. 10% Rule For each sample you want to load on a gel, make 10% more volume than needed...microliters can be lost in pipetting. For example, if you want to load 1.0 μg in 10 μL, make 1.1 μg in 11 μL. Reference...

  15. AAV ddPCR Titration

    Type
    Protocol
    ...droplet digital PCR (ddPCR) to titer purified recombinant Adeno-associated viral vectors (AAV). This protocol...ddPCR, read this blog post on ddPCR for AAV quantification. This protocol was modified from the publication...transfer the plate to the Droplet Reader. Open the QuantaSoft software to set up a new plate layout. Designate...

  17. Protocol - How to Perform Sequence Analysis

    Type
    Protocol
    ... Digest Introduction Sequence verification of important plasmid features (such as the gene/insert, fusion...you sequence any portion of the plasmid that is important for your experiments. There are a number of factors...

  18. Weighing Reagents Protocol

    Type
    Protocol
    ... or paper. You need to do this because you don’t want to include the weight of the weighing boat or paper...Conclusion Appropriately weighing reagents is important to ensure the accurate composition of your solutions...

  19. Video Library

    Type
    Protocol
    ...Over-Agar Antibiotic Plating Create plates to culture bacteria in the lab Over-Agar Antibiotic Plating ...

  20. Fluorescence Titering Assay

    Type
    Protocol
    ... It is not recommended to subject lentiviral supernatants to multiple freeze-thaw cycles. Procedure Seed...titer lentiviral vectors (10 5 TU/mL). If you anticipate that your viral stock will be higher titer, consider...
Showing: 1061 - 1080 of 1086 results