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We narrowed to 911 results for: TRO

Showing: 851 - 860 of 911 results
  1. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...of each well at 450 nm on the spectrophotometer. Use the spectrophotometer to generate a standard curve...Antibody Purification Protocol Antibody Guide Introduction This protocol describes how to run an indirect...antibody incubation and plate read Tips and Troubleshooting The optimal concentration of primary antibody...an automated ELISA plate washer. Equipment Spectrophotometer compatible with 96-well plates 1–10 µL single...Multichannel pipette reagent reservoirs Pipette controller Microcentrifuge Microplate shaker ELISA plate... antigen Purified recombinant antibody Isotype control antibody HRP-conjugated isotype-specific secondary...mix. Dilute the primary antibody and an isotype control antibody to the desired concentrations in the antibody...
  2. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...questions. Intro to the Lab Bench Basic Molecular Biology Plasmid Cloning Virus Antibodies Intro to the Lab... Lab Bench Introductory techniques designed to help you get started in the lab. Name Description (Link...Quantification Measure DNA concentration with a spectrophotometer Restriction Digests Cut and analyze plasmid...plasmid DNA with restriction enzymes Agarose Gel Electrophoresis Separate DNA by size on an agarose gel Watch...DNA using DNA ligase Bacterial Transformation Introduce DNA into a bacterial strain Watch the Video! CRISPR...Description Link to Video General Transfection Introduce plasmid DNA to mammalian cells Lentivirus Production.... Name Description Link to Video Transfection Introduce plasmid DNA to mammalian cells to produce antibodies...
  3. Protocol - Bacterial Transformation

    Type
    Protocol
    ...DNA Ligation Introduction Transformation is the process by which foreign DNA is introduced into a cell....should also add a positive control (many companies include a positive control plasmid with their competent...ligation of inserts to vectors than for an intact control plasmid. Incubate the competent cell/DNA mixture...transform large plasmids, it is a good idea to use electro-competent cells. Instead of relying on the heat-shock...heat-shock to cause the cells to take up the DNA, an electro-magnetic field is applied to the cell/DNA mixture... do this you will need to have access to an electroporator and the appropriate cuvettes. Follow the manufacturer's...
  4. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ...and FAQ Do controls When doing ligations you should ALWAYS do a vector alone + ligase control. This will...ligation. Additional controls are encouraged, but may only be required for troubleshooting failed ligations...The following table indicates the various controls: Control Ligase Interpretation Uncut vector - Checks...know has only been thawed once before. Always do controls. See Tips and FAQ below for details. Try different...treated, that the phosphatase treatment worked. This control should, in principle, be free of colonies, but ...
  5. DNA Quantification

    Type
    Protocol
    ...purity. This can be done with a spectrophotometer. A spectrophotometer uses the absorbance/transmission... instructions for the spectrophotometer specific to your lab. Spectrophotometer Tips Before measuring ...labs have a NanoDrop, which is a very small spectrophotometer that can accurately read DNA concentration...measuring any samples, be sure to ‘blank’ the spectrophotometer using the solution the DNA is resuspended in, ...NanoDrop. It is a good idea to re-zero any spectrophotometer between samples if they are expected to vary...
  6. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...an uncut sample of each vector to help with troubleshooting if your digests don’t look as you expected....product is the anticipated size and that your band is strong (indicating that the PCR reaction worked and that...insert. It is also important to set up negative controls in parallel. For instance, a ligation of the recipient...much less total DNA (<1ng) or if you are having trouble getting colonies, you might want to use higher ...to be very large (>10kb) you might want to use electro-competent cells instead of the more common chemically-competent...plasmid alone plate. The recipient plasmid alone control will tell you your “background” level or more specifically...see any colonies, you should conduct a positive control to ensure that your transformation worked. You ...
  7. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...Restriction Digest of Plasmid DNA Agarose Gel Electrophoresis Introduction Restriction enzymes are naturally occurring...and analyze the resulting fragments by gel electrophoresis . The pattern of the fragments on the gel can...to analyze a restriction digest. Equipment Electrophoresis chamber Pipetman Pipet tips Reagents Liquid...manufacturer's instructions) Gel loading dye Electrophoresis buffer Verifying Total Plasmid Size -OR- Insert...avoided. If you do have to do so, there is no way to control which orientation the insert is ligated into the... tricks see the Restriction Digest and Gel Electrophoresis pages....
  8. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ... uncut sample of each plasmid to help with troubleshooting if your digests don’t look as you expected....insert. It is also important to set up negative controls in parallel. For instance, a ligation of the recipient...much less total DNA (<1ng) or if you are having trouble getting colonies, you might want to use higher ...to be very large (>10kb) you might want to use electro-competent cells instead of the more common chemically-competent...plasmid alone plate. The recipient plasmid alone control will tell you your “background” level or more specifically...see any colonies, you should conduct a positive control to ensure that your transformation worked. You ...depending on the number of background colonies on your control plate (the more background, the more colonies you...
  9. Western Blot

    Type
    Protocol
    ...channel pipette Pipette controller Pipette tips and pipettes Spectrophotometer Heat block Mini gel tank...Protocol Recombinant Antibody Purification Protocol Introduction Western blot is a technique used to separate...ladder. Place the lid on the tank and plug the electrode cords into the power supply. Run the gel at 100... a dark room to detect the bands. Tips and Troubleshooting The optimal lysis buffer will vary depending...as expected and is specific, include a positive control sample that you know expresses the protein, such...transfected with the protein of interest, and a negative control sample that does not express the protein of interest...
  10. Lentivirus ddPCR Titration

    Type
    Protocol
    ...Viral Service Lentivirus Guide Virus Blog Posts Introduction This protocol describes how to use droplet digital...ribonuclease P/MRP 30 kDa subunit (RPP30) are used as a control for normalization. The dilution series outlined...wells containing the virus and the untransduced control. Pro-Tip For even seeding, prepare a batch for ...Determine the concentration of each sample on a spectrophotometer. Prepare 25 ng/µL stocks of each sample. Samples...calculate the final infectious titer. Tips and Troubleshooting We recommend wiping down all pipettes and equipment...concentration of RRE positive droplets in the untransduced control should be close to zero (A01). In this protocol...
Showing: 851 - 860 of 911 results