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

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We narrowed to 18 results for: PAN

Showing: 1 - 18 of 18 results

  1. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ... confluent, expand into larger vessels. A confluent well of a 6-well dish can be expanded into a 10 cm...reagent Day 3–14: Change media as needed Day 14–18: Expand and harvest stable cell lines Equipment Class II...undergo additional selection while the population expands. At this time, some researchers reduce the concentration...cm dish. A confluent 10 cm dish can be expanded into two 75 cm 2 flasks, etc. Pro-Tip This selection method...populations are growing well and have been sufficiently expanded, prepare cell stocks and/or harvest to test for...will have higher levels of expression. Consider expanding populations transduced with a variety of dilutions...Figure 1: Generation of monoclonal cell lines from expansion of individual A549 cells stably expressing Cas9...

  2. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ... Monitor cells for growth and expand cells Day 14–30: Analyze and expand monoclonal lines of interest ...cells have expanded but before they become over-confluent, trypsinize the cells and expand them to larger...After the monoclonal lines have been sufficiently expanded, screen the lines for transgene expression and...Figure 1: Generation of monoclonal cell lines from expansion of individual A549 cells stably expressing Cas9... in individual wells of a 96-well plate after expansion of single cells for 13 days. Note that growth ...Single cells from a stable cell pool were then expanded for an additional 3 weeks under blasticidin selection...stable cell pool (pool) and from individual cell expansions (monoclonal, 1-5). Wild-type A549 cells do not...

  3. Kit Free RNA Extraction

    Type
    Protocol
    ...). You can use either Isopropanol or Lithium Chloride for this step. Isopropanol (Option A): Add 1 volume...). You can use either Isopropanol or lithium Chloride for this step. Isopropanol (Option A): Add 1 volume...precipitation step, two options are also included: using Isopropanol or Lithium Chloride. This protocol was adapted...decontamination solution like RNase AWAY® or RNaseZAP® Isopropanol (for precipitation step, Option A) 7.5 M Lithium...volume of Isopropanol to the extracted aqueous layer. Incubate at -20°C for 1 hour. Lithium Chloride (Option...

  4. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...bacterial cultures , please see those pages. Several companies, such as (Link opens in a new window) NEB , (Link...phenol-chloroform Chloroform 100% ethanol or isopropanol 90% ethanol 70% ethanol TE buffer 3 M Na-acetate...plasmid-containing supernatant. Add either ethanol or isopropanol to precipitate the plasmid DNA. Either spin to...for 5 min. Note: Ribonuclease A (RNase A) is a pancreatic ribonuclease that digests single-stranded RNA...cold 100% ethanol or 350 μL room temperature isopropanol to the solution to precipitate the plasmid DNA...

  5. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...containing the trypan blue. Pipette 10 times to mix. Load 10 µL of the cell suspension/trypan blue mix into....45 µm PES filtering system, 500 mL, VWR 430770 Trypan Blue, Thermo Fisher T10282 Valproic acid sodium...flask 5–10 times before sampling. Transfer 10 µL of trypan blue into a clean microcentrifuge tube. Vortex ...

  7. Video Library

    Type
    Protocol
    ...positions he has held in his career, his favorite company tradition, and Addgene's role in the scientific... offers career advice for scientists looking to expand their international reach. Joel McDade Joel McDade...

  8. Protocol - Bacterial Transformation

    Type
    Protocol
    ...commonly referred to as 'competent cells.' Many companies sell competent cells, which come frozen and are... You should also add a positive control (many companies include a positive control plasmid with their ...

  9. Lentivirus Production

    Type
    Protocol
    ...without limiting cell growth. Left panels: bright field images; right panels: GFP channel images. Last reviewed...

  10. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...985 (Link opens in a new window) . Figure 2: Left panel: Iodixanol gradient after ultracentrifugation. The...): 973-985 (Link opens in a new window) . Right panel: cartoon indicating the position of the needle for...

  11. AAV Production in HEK293 Cells

    Type
    Protocol
    ...until no live cells can be seen when stained with Trypan Blue. Pellet cell debris by centrifugation at 3900...started with cells that are ~80% confluent (center panel). These cells were imaged with a 10X objective and...

  14. Centrifugation

    Type
    Protocol
    ... with microfuge tubes in place, and the control panel where you can set the time and speed needed for ...

  16. General Transfection

    Type
    Protocol
    ...fluorescent protein expression (green in the right panels) with a limited effect on cell growth. Last reviewed...

  18. CRISPR Library Amplification

    Type
    Protocol
    ...electroporation. Arcing would manifest as a loud pop often accompanied by some light in the cuvette during the electroporation...
Showing: 1 - 18 of 18 results