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Showing: 141 - 160 of 165 results
  1. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ..., VWR 89130-902 50 mL conical tubes, VWR 89039-656 Microcentrifuge tubes, VWR 89126-724 Filtering system...fractions into the tubes containing 50 µL 1 M Trizma hydrochloride pH 9.0. Cap the tubes and vortex for 5...from the collection tube each time. Place column in a new 50 mL LoBind collection tube. Slowly apply the...Benchtop centrifuge compatible with 50 mL conical tubes NanoDrop spectrophotometer 37 °C, 5% CO 2 incubator... column, Millipore Sigma UFC903024 50 mL LoBind tubes, VWR 76289-498 Before Starting Wipe down all pipettes.... re Collect the flow-through in a 50 mL conical tube and dispose after use. Carefully pour the filtered...Trizma hydrochloride pH 9.0 to 10 microcentrifuge tubes. Cap the Gravitrap columns at the bottom. Add 5 ...
  2. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...mix to add to each PCR tube. Secure the tops to the PCR tubes, gently tap each tube to bring all the liquid...times. Keep tube on ice. Add the forward and reverse primers to the thinned walled PCR tubes. Note: Do ...List Reagents for each 50µL PCR: Thin-walled PCR tube Ice Bucket 2 μL Template DNA (10 ng-500 ng) 5 μl...resource for choosing primers. Place thin-walled PCR tubes on ice Set up a 50 μL reaction (Keep all your reagents...DO NOT get added to a master mix. Place reaction tubes in PCR machine. Set annealing temperature 5°C below... we multiply each volume by 7. In a single 1.5mL tube combine the following: 10X Taq buffer with MgCl ...added, pipet the forward primer onto one side of the tube wall and the reverse primer onto the other. Add ...
  3. Lentivirus ddPCR Titration

    Type
    Protocol
    ...PCR tube strip into a chilled 96-well freezer block. Add 16 µL of the master mix to each PCR tube. Be ...dispense to the bottom of the tube without collecting drops along the side of the tube. Add 4 µL of the 25 ng... cartridge holder, Bio-Rad, 1863051 8-strip PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, ...Polystyrene reservoirs, VWR, 89094-662 Microcentrifuge tubes, VWR, 87003-294 Primers/probe targeting RRE: forward...virus in DMEM complete: Prepare 7 microcentrifuge tubes with the appropriate amount of media volume listed... DMEM complete and transfer to a microcentrifuge tube. Centrifuge for 5 min at 100 x g . Gently aspirate...Prepare the RRE/RPP30 master mix in a microcentrifuge tube as shown below. For 8 samples prepare enough master...
  4. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...hours. c. In polypropylene microfuge tubes (do NOT use polystyrene tubes), make a cocktail for each transfection...walls of the tube before it has been diluted. Mix by swirling or gently flicking the tube. Incubate for...not onto the walls of the tube. Mix by swirling or gently flicking the tube. f. Incubate for 20-30 minutes... 7kb band and place in a sterile microcentrifuge tube. TIP: When visualizing DNA fragments to be used ...Streptomycin Invitrogen: #15140-122 Polypropylene tubes VWR: #87003-290 Note: pLKO.1 could also be packaged... μL FuGENE® + 740 μL OPTI-MEM In a polypropylene tube, add OPTI-MEM first. Pipette FuGENE® directly into...temperature. e. Add 80 μL of FuGENE® master mix to each tube from step c for a total volume of 100 μL. Pipette...
  5. Protocol - How to Create a Bacterial Glycerol Stock

    Type
    Protocol
    ... lid and the tube of a glycerol stock before you place the sample at -80°C. Frozen tubes are hard to write...culture to 500 μL of 50% glycerol in a 2 mL screw top tube or cryovial and gently mix. Notes: Make the 50% ...solution by diluting 100% glycerol in dH20. Snap top tubes are not recommended as they can open unexpectedly...unexpectedly at -80°C. Freeze the glycerol stock tube at -80°C. The stock is now stable for years, as long as it...recover bacteria from your glycerol stock, open the tube and use a sterile loop, toothpick or pipette tip...for long periods at -80°C can lose labels stuck to tube! Reference Page Top Index...
  6. Protocol - Bacterial Transformation

    Type
    Protocol
    ...microcentrifuge or falcon tube. GENTLY mix by flicking the bottom of the tube with your finger a few times...shock each transformation tube by placing the bottom 1/2 to 2/3 of the tube into a 42°C water bath for...42 °C Ice bucket filled with ice Microcentrifuge tubes Sterile spreading device Reagents LB agar plate ... on the competent cells you are using). Put the tubes back on ice for 2 min. Add 250-1,000 μl LB or SOC...transformation efficiency (such as when you have a tube of plasmid DNA and just need to transform bacteria...
  7. Lentivirus Production

    Type
    Protocol
    ...Microcentrifuge tubes, Neptune 3745.X 10 cm tissue culture dish, Corning 430167 15 mL conical tubes, VWR 21008...membrane. Aliquot 500–1000 μL into sterile tubes. Store the tubes at -20 °C or -80 °C. After thawing, the... for up to 2 months. After 2 months, discard the tube and thaw a new working stock. The optimal mass DNA...PEI dropwise while gently flicking the diluted DNA tube. Incubate the mixture 12–15 min at RT. During the...transfer the harvested media to a polypropylene storage tube and store at 4 °C between harvest. Centrifuge the...
  8. Protocol - How to Purify DNA from an Agarose Gel

    Type
    Protocol
    ...Place the gel in a labeled microfuge tube. Using a scale, weigh the tube with the gel fragment after zeroing... with an empty tube. Alternatively, you can just subtract the weight of the empty tube from the weight...weight of the tube with the gel fragment. The weight of the gel is directly proportional to its liquid volume...
  9. General Transfection

    Type
    Protocol
    ...membrane. Aliquot 500–1000 µL into sterile tubes. Store the tubes at -80 °C. After thawing the solution can...Polyethylenimine, linear MW 25,000 Da Microcentrifuge tubes, Neptune 3745.X 10 cm tissue culture dish, Corning... for up to 2 months. After 2 months, discard the tube and thaw a new working stock. Considerations Before...PEI dropwise while gently flicking the diluted DNA tube. Incubate the mixture 15–20 min at RT. Carefully...
  10. Pipetting Protocol

    Type
    Protocol
    ...microcentrifuge tube, a bottle etc.). If there is already other liquid in the receiving tube, place the pipette...Containers to hold measured liquid (ex: microfuge tube, bottle, etc.) Labels for containers Reagents Liquid... avoid touching the pipette to the inside of the tube. Only the tip should be in the liquid. Pro-Tip If...pipetting the same amount of liquid into different tubes or into wells of a microplate, you may consider ...
  11. Weighing Reagents Protocol

    Type
    Protocol
    ...microcentrifuge tube on the balance and then tare it. Then add the material directly to the tube to weigh it...paper Balance Scoopula or spatula Container (beaker, tube, flask, etc.) Reagents Material that will be weighed...They also help you transfer the material to your tube or container. Pro-Tip If you’re using weighing paper...
  12. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ... into individual tubes and then add the appropriate amount of Master Mix to each tube. This will save ... double digest finder tool from NEB . In a 1.5mL tube combine the following: DNA Restriction Enzyme(s)...usually better. Mix gently by pipetting. Incubate tube at appropriate temperature (usually 37 °C) for 1...
  13. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ...good idea to take a fresh tube, thaw it one time and aliquot individual tubes of 5, 10 or 20μL for storage...use. Combine the following in a PCR or Eppendorf tube: Vector DNA Insert DNA Ligase Buffer (1μL/10μL reaction...set up ligations in the future you can thaw a new tube that you know has only been thawed once before. ...
  14. AAV Production in HEK293 Cells

    Type
    Protocol
    ...membrane. Aliquot 500–1000 μL into sterile tubes. Store the tubes at -80 °C. After thawing, the solution ... for up to 2 months. After 2 months, discard the tube and thaw a new working stock. 40% POLYETHYLENE GLYCOL... min at 4 °C. Transfer the cleared lysate to the tube containing the resuspended virus from step 12 above...
  15. Immunocytochemistry

    Type
    Protocol
    ...medium Laboratory wipes 15 mL conical tubes 50 mL conical tubes Before Starting Refer to the manufacturer's...channel pipette Reagents 1X PBS Microcentrifuge tubes Sterile Poly-D-lysine coated coverslips HeLa cells...
  16. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...Microcentrifuge tubes, Neptune 3745.X 6-well tissue culture treated dish, Corning 3516 15 mL conical tubes, VWR ...polybrene. Mix well by pipetting or inverting the tube. Aliquot 1 mL of cell suspension (i.e., 50,000 cells...
  17. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ... Gel loading tips, Corning 4853 Microcentrifuge tubes, Neptune 3745.X PBS, 1X pH 7.4, VWR 45000-446 250...PBS Transfer 5 µL of each standard to a microfuge tube. Add 8 µL of PBS to a total volume of 13 µL . Prepare...Transfer 5 µL of recombinant antibody to a microfuge tube. Add 8 µL of PBS to total 13 µL . Prepare 1X MOPS...
  18. Western Blot

    Type
    Protocol
    ... buffer e.g., RIPA lysis buffer Microcentrifuge tubes BCA Assay, Thermo Fisher 23227 β-mercaptoethanol...1 mL of 1X PBS and transfer to a microcentrifuge tube. Centrifuge for 5 min at 100 x g . Carefully remove... Transfer supernatant to a clean microcentrifuge tube. Use the lysate immediately or store at -80 °C until...
  19. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...When ready to grow your culture, add liquid LB to a tube or flask and add the appropriate antibiotic to the... you will usually want to start 2 mL in a falcon tube, but for larger preps you might want to use as much...
Showing: 141 - 160 of 165 results