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Addgene
Showing: 81 - 100 of 108 results
  1. AAV Production in HEK293 Cells

    Type
    Protocol
    ...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Hemocytometer or cell counter Ice bucket CO 2 incubator Pipet controller Hazardous...Neutralize trypsin by adding 10 mL of DMEM Complete. Pipet back and forth vigorously multiple times to obtain...with the cells harvested in the previous step. Pipette back and forth or shake vigorously to obtain a ...total of 5 mL of cell lysis buffer (recipe above). Pipet back and forth to resuspend each pellet completely...adding a total of 12.5 mL of cell lysis buffer. Pipet back and forth to resuspend each pellet completely...
  2. Immunocytochemistry

    Type
    Protocol
    ...Fix and label cells Equipment Pipette controller Pipette tips and pipettes Rocking platform Tweezers Fluorescent...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Reagents 1X PBS Microcentrifuge tubes Sterile Poly-D-lysine coated coverslips HeLa...
  3. Western Blot

    Type
    Protocol
    ...single channel pipette Pipette controller Pipette tips and pipettes Spectrophotometer Heat block Mini gel...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...
  4. Fluorescence Titering Assay

    Type
    Protocol
    ...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Inverted fluorescence...cells into 14 mL of DMEM complete. Mix well by pipetting or inverting. Aliquot 2 mL of cell suspension ...
  5. Lentivirus Production

    Type
    Protocol
    ...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container pH meter Stir...out of the previously seeded 10 cm plate. Slowly pipette the transfection mix onto the 10 cm plate, being...
  6. Colony Formation Titering Assay

    Type
    Protocol
    ...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents DMEM...complete containing 10 μg/mL polybrene. Mix well by pipetting or inverting. Aliquot 1.35 mL of the cell suspension...
  7. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...Biosafety Cabinet 4 °C Refrigerator Micropipettes and tips Pipettor and pipettes Benchtop centrifuge compatible...Reagent Preparation instructions. Wipe down all pipettes and equipment with 10% bleach prior to use. Reagent...Check the cell density and viability: Using a 5 mL pipette, transfer 0.5 mL of HEK293 cell suspension into...the microfuge tube containing the trypan blue. Pipette 10 times to mix. Load 10 µL of the cell suspension...
  8. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents DMEM...DMEM complete + 10 µg/mL polybrene. Mix well by pipetting or inverting the tube. Aliquot 1 mL of cell suspension...
  9. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette CO 2 incubator Pipet controller Hazardous waste container Microscope Hemocytometer... by passing several times through a serological pipet or by passing through a 40 µm cell strainer mesh...
  10. Kit Free RNA Extraction

    Type
    Protocol
    ...storage in the cold room may damage it. Using a pipettor, transfer the top, aqueous phase to a new RNAse-free...disturb or collect the interphase layer with your pipette tip. Pro-Tip To collect as much of the top aqueous...interphase layer, consider using a lower volume pipettor like a p200 to collect a majority of the aqueous...
  11. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ...spread antibiotic over-agar. Equipment Pipette tips for both pipetting and spreading Bunsen burner (or other...diameter petri plate containing solidified LB-agar, pipette 150 µL of carbenicillin on top of the agar and ...liquid remaining on the surface). We use the micropipette tip itself to do the spreading; the tip is gently...same way as the antibiotic, using either a bent micropipette tip or other cell spreading device that fits...
  12. General Transfection

    Type
    Protocol
    ...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents DMEM...
  13. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...is unloaded, push the pipettor to the second stop and carefully raise the pipette straight out of the buffer...comb or towards the sides/edges of the gel with a pipette tip. Place newly poured gel at 4 °C for 10-15 mins... the tip. Place the very top of the tip of the pipette into the buffer just above the well. Very slowly...needed because several microliters can be lost in pipetting. For example, if you want to load 1.0 μg in 10...
  14. CRISPR Library Amplification

    Type
    Protocol
    ...Spreaders (VWR, 60828-680) 5 mL and 10 mL Serological pipettes Ice slurry (Ice bucket with ice and water to create...until completely thawed. Chill a box of 200 µL micropipette tips in a -20 °C freezer. Aliquot 3 mL SOC into... two separate transformations). Pro-Tip Do not pipette repeatedly or mix when removing SOC containing ... Use one scraper for all plates. Use two 10 mL pipettes (one for dispensing cold LB and one for removing...). Pro-Tip Scraped bacteria in LB can clog the pipette either mix gently up and down avoiding introducing...
  15. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...be worn at all times. Use plastic pipettes in place of glass pipettes or needles. Liquid waste should be...OPTI-MEM In a polypropylene tube, add OPTI-MEM first. Pipette FuGENE® directly into the OPTI-MEM – do not allow...tube from step c for a total volume of 100 μL. Pipette master mix directly into the liquid and not onto...cells. Swirl to disperse mixture evenly. Do not pipette or swirl too vigorously, as you do not want to ...fresh DMEM + 10% FBS + penicillin/streptomycin. Pipette the media onto the side of the plate so as not ...
  16. Video Library

    Type
    Protocol
    ...Mulitchannel Pipetting Technique Learn how to pipette multiple samples at once using a multichannel pipette and...and avoid inconsistent pipetting with our simple tips Bacterial Transformations We walk you through a standard...
  17. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ...digest results. Equipment Electrophoresis chamber Pipetman Reagents Liquid DNA aliquot of your plasmid of...instructions) Gel loading dye Electrophoresis buffer Pipet tips Procedure Select restriction enzymes to digest....2-0.5 µL of enzyme because it is difficult to pipette less volume than this; 0.2-0.5 µL will likely be... more enzyme is usually better. Mix gently by pipetting. Incubate tube at appropriate temperature (usually...
  18. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...minimizes the number of small volumes you need to pipet. If you are using the same template DNA for all ...wish to perform and add 10% extra to account for pipetting error. In this example, we have 7 different reactions... (0.14µl) = 1.54µl Mix the contents by gently pipetting up and down several times. Keep tube on ice. Add...mix so you know that the primers have been added, pipet the forward primer onto one side of the tube wall...
  19. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...chromosomes. Collect the supernatant into a new tube by pipetting or carefully pouring. Optional: Add 5 μL of 2 ...particles Bottom phase - Organic phase (protein) Pipet the aqueous DNA layer and place it in a new microfuge...the tube after the 70% ethanol wash. You can also pipet the supernatant out of the tube if you are worried...
Showing: 81 - 100 of 108 results