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

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We narrowed to 1,018 results for: PLE;

Showing: 961 - 980 of 1018 results

  10. Lentivirus ddPCR Titration

    Type
    Protocol
    ...layout is complete, select 'Run' to begin the droplet reading. When the droplet reading is complete, export...concentration of each sample on a spectrophotometer. Prepare 25 ng/µL stocks of each sample. Samples can be used...the droplet generator. Close the lid and wait for the droplets to be generated. Once the droplets have...the droplets. Transfer the droplets to a prechilled PCR plate. Pro-Tip To ensure that the droplets are...constant across samples. In the RRE example below, 2-fold serial dilutions of a sample were loaded in ...Lentivirus sample data, RPP30 positive (green) and negative (black) droplets Sample ID Plasmid ID Dilution...incubating with 200 µL TrypLE for 1–2 min. Resuspend cells in 500 µL DMEM complete and transfer to a microcentrifuge...

  11. AAV ddPCR Titration

    Type
    Protocol
    ...layout is complete, select 'Run' to begin the droplet reading. When the droplet reading is complete, export...the droplet generator. Close the lid and wait for the droplets to be generated. Once the droplets have...the droplets. Transfer the droplets to a prechilled PCR plate. Pro-Tip To ensure that the droplets are... ddPCR sample data Sample Dilution Concentration Titer (GC/mL) Average Titer (GC/mL) AAV Sample 1 6400000... used for master mix preparation and droplet generation (droplet generation BSC) to ensure no contamination...tube as shown below. For 8 samples prepare enough master mix for 9 samples. ITR Master Mix Volume 9X Volume... the PCR tubes. Generate the Droplets Bring the PCR tubes to the droplet generation BSC. Without disturbing...

  12. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...empty NTC C AAV reference Sample 3 D E Sample 1 Sample 4 F E Sample 2 Sample 5 F Perform data analysis...most samples. If sample is expected to have a titer <1 x 10 12 GC/mL, use dilutions 3–6 If sample is expected...between them. Sample Data Figure 1: Example of a valid 8-point standard curve. Figure 2: Example of the amplification...standards and samples in duplicate at least If possible, include an AAV reference sample of known titer...error and variability Mix samples very well by pipetting back and forth multiple times at each step Reagent...Count the number of samples (n) and prepare master mix for an additional 10 samples (n+10 – the additional...that there is enough master mix for all samples). Each sample requires 15 μL of master mix. Pro-Tips Use...

  13. Kit Free RNA Extraction

    Type
    Protocol
    ... 10 seconds. Incubate sample(s) for 15 minutes on ice and centrifuge the sample(s) for 15 minutes at 12,000...RNA. Store your RNA sample(s) at -80 °C to prevent RNA degradation and avoid multiple freeze-thaw cycles...cycles. Pro-Tip To avoid multiple freeze-thaw cycles of your entire RNA sample, consider making smaller aliquots...RNA.Store your RNA sample(s) at -80 °C to prevent RNA degradation and avoid multiple freeze-thaw cycles...Always wear gloves, and whenever possible, keep RNA sample and reagents cold and work quickly to reduce RNA...use 1 mL of Solution D per 1 X 10 7 cells. Allow sample(s) to sit at room temperature for 5 minutes to ...

  14. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...of 4X sample buffer to each sample. Add 2 µL 10X reducing agent to each sample. Spin the sample briefly...concentration of your samples based on your sample’s HC + LC area. Example calculation: If the sample HC + LC Area...Invitrogen NP0001 4X NuPage sample buffer, Invitrogen NP0007 10X NuPage sample reducing agent (500 mM dithiothreitol...microcentrifuge. Heat the samples for 10 min at 100 °C in a heat block. Spin the sample briefly in the microcentrifuge...one lane before loading samples. Load 20 µL of each recombinant antibody sample to the appropriate well... mobility of the bands (sample AR0016 in Figure 1) indicates that the samples may not have been processed...area of all bands in the lane Example for AR0018 (lane 2 in Figure 1): Sample Peak 1 (contaminant) Peak 2...

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

    Type
    Protocol
    ...binding capacity. If your sample exceeds the capacity, divide the sample among multiple columns. For information...4 mL of recombinant antibody sample. Pro-Tip If the volume of the sample is greater than 4 mL , then divide... Slowly apply the sample to the center of the compact resin bed. Pro-Tip For sample volumes <1.5 mL top...the sample with 0.2 mL PBS to aid protein recovery. Allow the resin bed to fully absorb the sample. Centrifuge...Spectrophotometer to see if the sample has reached the desired concentration. If the sample concentration is still... to see if the sample has reached the desired concentration. Gently transfer the sample from the reservoir...similar outcomes when using these protocols. However, please be aware that the protocol may need to be adjusted...

  16. Using a Light Microscope Protocol

    Type
    Protocol
    ...helps concentrate the light onto the sample, and then through the sample. However, some of that light won’...hitting the sample or b) using dyes or stains that add color to some components of the sample but not others... power objective to find your sample. Place your slide (or other sample type) on the microscope stage....so that the light shines onto your sample. To ensure that your sample is adequately illuminated, look through... coarse focus knob to bring your sample into focus. Once your sample is in focus, use the stage arm to... you can easily see your sample. Then use the focus knobs to bring the sample into focus. Pro-Tip As you...magnification used, any stains used in your sample, and sample information so that you or others can interpret...

  17. Centrifugation

    Type
    Protocol
    ... which you need to spin your samples. For example, spinning down samples in 1.5 mL microfuge tubes at ...liquid sample. It achieves this by using centrifugal force, which is generated by spinning the sample at ... with water to the same volume as your samples. If your samples are significantly higher density than ...centrifuge to separate different components in a liquid sample. Protocols... containers, spin at different speeds, or keep samples at specific temperatures. This protocol will cover...for the biosafety level of the lab space Reagents Sample to be centrifuged Water Background Information ...speed needed for your experiment. Procedure An example of a balanced tabletop centrifuge, with tubes evenly...

  18. Western Blot

    Type
    Protocol
    ...duplicate samples on the plate. Subtract the average absorbance of the blank from all of the samples. Plot...concentration of the sample from the standard curve. Determine the volume of sample required to load an...protein for each sample. Pro-Tip The ideal total protein loaded will vary between samples and target proteins...abundance in the sample you will need to load a greater amount of total protein. Prepare sample for loading...Dilute 4X protein loading buffer in the sample to 1X . Boil the samples for 10 min at 100 °C . Section 3: SDS-PAGE...similar outcomes when using these protocols. However, please be aware that the protocol may need to be adjusted...blotting to visualize a protein from cells or tissue samples. Equipment Microcentrifuge 0.5–10 µL single channel...

  19. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...steadily, push the sample out and watch as the sample fills the well. After all of the sample is unloaded, .... Loading Samples and Running an Agarose Gel: Add loading buffer to each of your DNA samples and mix well...Note: When loading the sample in the well, maintain positive pressure on the sample to prevent bubbles or...your sample lanes. For more details on doing diagnostic digests and how to interpret them please see the...). Microwave for 1-3 min until the agarose is completely dissolved (but do not overboil the solution, ...the final percentage of agarose in the gel. Many people prefer to microwave in pulses, swirling the flask...room temperature for 20-30 mins, until it has completely solidified. Pro-Tip If you are in a hurry, the...

  20. AAV Production in HEK293 Cells

    Type
    Protocol
    ...trypsin by adding 10 mL of DMEM Complete. Pipet back and forth vigorously multiple times to obtain a single ...Thermo Fisher 31985-070 0.05% Trypsin/EDTA (e.g. TrypLE, Thermo Fisher, 12605010) 1X PBS pH 7.4 without... mM NaCl, 2 mM MgCl2) Reagent Preparation DMEM Complete : 10% v/v FBS and 4 mM L-alanyl-L-glutamine (or...Chloride Close the bottle and mix by inverting multiple times Adjust pH to ~8.5 Filter sterilize through...-175 flasks. Adjust volume to 300 mL with DMEM complete media and mix. Seed all cells in 1 CS2. Return...CS2 to help detach the cells, add 200 mL of DMEM complete media, then transfer the cells into a sterile ...sterile bottle. Rinse the CS2 with 100 mL of DMEM complete medium and pool with the cells harvested in the previous...
Showing: 961 - 980 of 1018 results