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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 25 results for: erf

Showing: 1 - 20 of 25 results

  1. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...Protocol for how to perform a diagnostic digest to verify plasmid size, verify insert orientation, and...expected sizes. This is frequently done after performing either PCR - or restriction enzyme -based cloning...

  3. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...your purified virus. Nonetheless, we recommend performing a buffer exchange before using the purified AAV... carefully puncture the QuickSeal tube at the interface of the 60% and 40% gradient (see Figure 2) with...Avoid the proteinaceous material near the 40–25% interface. Pro-Tip As soon as the top of the tube is punctured...solution will flow from the needle set at the 40–60% interface. Make sure that the microcentrifuge tubes are ... the QuickSeal tube slightly below the 60–40% interface with an 18 ga needle attached to a 10 mL syringe...collecting the proteinaceous material at the 40–25% interface. Repeat for each QuickSeal tube. (Optional) For...ultracentrifugation. The arrow indicates the 60–40% interface. The vertical black line indicates the location...

  4. CRISPR Library Amplification

    Type
    Protocol
    ...Follow this protocol to perform amplification of CRISPR pooled plasmid libraries in Escherichia coli ...Estimated time 2-3 hours) Transformation should be performed at the end of the day to ensure that growth times...shaking period, pool and gently mix the four tubes. Perform sequential 1:100 dilutions of the cells (add 10...10 µL of the pool to 990 µL LB then perform a second and then third 1:100 dilution). Plate 100 µL of each... recombined fraction but care must be taken to perform adequate NGS based QC to ensure no change in representation...to the pre-amplified stock. A last option is to perform a gel extraction of either the original sample ...of the DNA, but please note that NGS should be performed to ensure representation is maintained. Maxipreps...

  5. Ligation Independent Cloning

    Type
    Protocol
    ...standard polymerase protocol), causing the enzyme to perform exonuclease activity until the first "G" in the...sterile dH20 to 40 μl Step 4: Amplify Insert by PCR Perform PCR amplification of your insert following the ...your PCR product before proceeding, as they will interfere with the exonuclease activity of T4 Pol in the...

  6. Using a Light Microscope Protocol

    Type
    Protocol
    ...phone-sized, foldable microscopes that aren't very powerful (but are cheap and accessible) to massive transmission... are multiple objective lenses. Each objective performs essentially the same task but has different magnifying...scratches, and dust can easily impair microscope performance. Be sure to clean up your work area and microscope...

  7. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ... follow the kit's instructions. If you want to perform plasmid purification without using a kit, you can...ribonuclease that digests single-stranded RNA. Optional: Perform phenol-chloroform extraction - see protocol below... DNA purified without a kit it is advisable to perform a phenol/chloroform extraction of the supernatant...

  8. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...Watch the protocol video below to learn how to perform Polymerase Chain Reaction (PCR). Basic PCR Program...pairs. The initial denaturation step is commonly performed at 94–98°C. Denature 30 seconds at 94°C: Continued...reagent by the number of individual PCRs you wish to perform and add 10% extra to account for pipetting error...

  9. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ... of a stable cell culture after transduction. Performing antibiotic selection on transduced cells enables...5 dilutions plus one 'no virus' control well). Perform a "reverse transduction" by seeding 50,000 cells...untransduced well (0 µL lentivirus, above) are dying. Perform regular media changes and monitor the growth of...

  10. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...monoclonal cell lines. When this protocol was performed at Addgene on the A549 cells described in our ...expression and/or other phenotypes. For example, perform Western blotting to screen for lines with the highest...blasticidin selection. Anti-Cas9 Western blotting was performed on whole-cell extracts from the initial stable...

  12. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ...fragments. This reaction, called ligation, is performed by the T4 DNA ligase enzyme. The DNA ligase catalyzes...digested vector backbone with a phosphatase before performing the ligation reaction (phosphatase removes the...

  13. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...Standard protocol for performing agarose gel electrophoresis, including tips to improve resolution and...Watch the protocol video below to learn how to perform gel electrophoresis. Equipment Casting tray Well...

  16. General Transfection

    Type
    Protocol
    ... (am): Observe fluorescence, harvest cells, or perform your experiment Equipment Class II, Type A2 Biological...

  18. Pipetting Protocol

    Type
    Protocol
    ...the Volume on the Pipette Although each pipette performs the same function, the numbers are read differently...

  20. Pouring LB Agar Plates

    Type
    Protocol
    ...technique if you are working with any weird and wonderful organisms. While your samples are sterilizing ...
Showing: 1 - 20 of 25 results