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Addgene

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

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We narrowed to 30 results for: SET

Showing: 1 - 30 of 30 results

  1. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...remove the plate seal from the 96-well plate and set aside. Aspirate the wells and use a multichannel ...the plate. Place the plate on a microplate shaker set at 400 rpm and shake for 1 min . Repeat steps 8–10...a plate seal and incubate on a microplate shaker set to 400 rpm for 2 h at room temperature or overnight...the plate Place the plate on a microplate shaker set at 400 rpm and shake for 1 min . Repeat steps 18–...

  2. Water Bath Protocol

    Type
    Protocol
    ...water bath on and set the appropriate temperature as described in your protocol. How to set the temperature...dial or a digital display you can set to the correct temperature. Set up your water bath so that it will...also helps the water bath heat up quicker. Pro-Tip Set up the water bath 30 minutes to 1 hour before you...

  3. Pipetting Protocol

    Type
    Protocol
    ...Without moving past the maximum possible setting on the pipette, set the desired volume of the liquid on the...tip is properly set on the pipette to ensure pipetting accuracy. Once the pipette is set and you have the... pipette tip, the volume display which shows the set volume for the the pipette, and the tip ejector ring...

  4. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...PCR machine. Set annealing temperature 5°C below the primer melting temperature (Tm). Set extension step...create thousands of identical copies using a simple set of reagents and a basic heating and cooling (denaturing...choosing primers. Place thin-walled PCR tubes on ice Set up a 50 μL reaction (Keep all your reagents on ice...designed to have similar melting temperatures (Tm). Set the annealing temperature to 5°C lower than the Tm...MgCl 2 and/or 1µl DMSO to each reaction. The ideal setup for this troubleshooting step is to do one reaction...

  5. AAV ddPCR Titration

    Type
    Protocol
    ...multichannel pipette set to 5 µL. For dilutions 6–8, use the 20–200 µL multichannel pipette set to 50 µL. For...dilutions, use the 20–200 µL multichannel pipette set to 90 µL and mix by pipetting the liquid up and down...ddPCR plate onto a chilled 96-well freezer block and set aside in the droplet generation BSC to cool. Prepare... Droplet Reader. Open the QuantaSoft software to set up a new plate layout. Designate the sample name,...

  6. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...the recipient plasmid, it is useful to see if one set of enzymes will work in the same restriction enzyme...future steps. Experimental Procedure Digest your DNA Set up restriction digests for your donor and recipient...recipient plasmid to insert. It is also important to set up negative controls in parallel. For instance, a...

  7. Centrifugation

    Type
    Protocol
    ...tubes in place, and the control panel where you can set the time and speed needed for your experiment. Procedure... close the external lid. Adjust the centrifuge settings according to speed and time (and temperature, ...

  8. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ...: Standard Insert + Vector DNA Ligation Before setting up the ligation reaction itself, it is important... 20μL for storage at -20°C. Whenever you need to set up ligations in the future you can thaw a new tube...

  9. Ligation Independent Cloning

    Type
    Protocol
    ... no additional water (<5 μl). It is advisable to set up multiple reactions by holding the vector concentration...insert, thereby increasing your chances of success. Set up a vector only control with water instead of the...

  10. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...Estimate of time required: ~3 h Workflow Timeline Plate set-up: 2 h qPCR run: 1.5 h Data analysis: 30 min Equipment...load the standards and samples onto the qPCR plate Set up and load the 96-well plate: Load 5 μL of each ...repeat 39x from step 3 / melt curve Example of plate set-up: 1 2 3 4 5 6 7 8 A 1.00 x 10 9 1.00 x 10 8 1.00...

  11. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...spectrophotometer 37 °C, 5% CO 2 incubator with shaking platform set to 120 rpm 37 °C bead bath Clamp stand and clamps... NanoDrop Spectrophotometer using the A280 IgG setting. Combine all eluates with measurable protein. Determine... NanoDrop Spectrophotometer using the A280 IgG setting. If the concentration of the pooled sample is above...

  12. Lentivirus ddPCR Titration

    Type
    Protocol
    ...remove virus from pipette tip. To mix the dilution, set the P200 pipette to 200 µL and pipette 10–20 times...ddPCR plate onto a chilled 96-well freezer block and set aside in the BSC to cool. Prepare the RRE/RPP30 master... Droplet Reader. Open the QuantaSoft software to set up a new plate layout. Designate the sample name,...calculate the titer. Calculations In the experimental setup above, the following virus dilutions were used to...factor v = Virus volume, mL *In the experimental setup above, the cells per well is 300,000 and the virus...

  13. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...counter 37 °C, 5% CO 2 incubator with shaking platform set to 120 rpm 37 °C bead bath Vortex Stir bar Magnetic...a 37 °C, 5% CO 2 incubator on a shaking platform set to 120 rpm. Pro-Tip Do not use cells that are over...into a clean microcentrifuge tube. Pro-Tip Cells settle quickly and need to be resuspended before sampling...

  14. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...ready for restriction digestion. Digest Your DNA Set up restriction digests for your PCR product and recipient...recipient plasmid to insert. It is also important to set up negative controls in parallel. For instance, a...

  15. Protocol - How to Streak a Plate

    Type
    Protocol
    ...and your initials. Labeling within a laboratory setting is important for organization, and it is recommended...

  16. Weighing Reagents Protocol

    Type
    Protocol
    ...measurement. Hit the tare button (or zero button) to set the balance to zero. Pro-Tip If you’re weighing a...

  18. Video Library

    Type
    Protocol
    ...technique, including personal protective equipment, setting up a clean workspace, and maintaining sterility...

  20. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...solidified. Pro-Tip If you are in a hurry, the gel will set more quickly if you place the gel tray at 4 °C earlier...increases the density of your DNA sample causing it settle to the bottom of the gel well, instead of diffusing...

  22. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...AAV-containing Iodixanol solution will flow from the needle set at the 40–60% interface. Make sure that the microcentrifuge...Protocol Video Watch the protocol video below to learn setup and use an iodixanol column gradient for AAV purification...forth a few times to mix the iodixanol that has settled at the bottom of the column. We recommend concentrating...

  23. Western Blot

    Type
    Protocol
    ...follows: Unseal the iBlot 2 PVDF Mini transfer stack. Set the Top Stack to one side and discard the white separator...

  24. Pouring LB Agar Plates

    Type
    Protocol
    ...Place the gel mix in the autoclave and run on a setting that gets the sample to at least 121 ℃ under 20...

  25. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...selection. Use an siRNA selection tool to determine a set of top-scoring targets for your gene. For example...help guide your reaction buffer selection when setting up double digests. Digest pLKO.1 TRC-cloning vector...

  26. CRISPR Library Amplification

    Type
    Protocol
    ...2023 Workflow Timeline Day 1: Transform, recover, set up overnight growth (Estimated time 2-3 hours) Transformation...

  27. General Transfection

    Type
    Protocol
    ... transfected using 1:1, 1:2, 1:3 and 1:6 µg of pRosetta :µg of PEI. The 1:2 and 1:3 ratios provided high...

  28. Lentivirus Production

    Type
    Protocol
    ...were transfected with the GFP-expression plasmid pRosetta using μg total DNA to μg PEI ratios of 1:1, 1:...

  29. Colony Formation Titering Assay

    Type
    Protocol
    ...indicated serial dilutions of the lentiviral vector pRosetta . The following day, cells were treated with puromycin...
Showing: 1 - 30 of 30 results