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Addgene

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

In Category

We narrowed to 17 results for: USP

Showing: 1 - 17 of 17 results

  1. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...20 min. Resuspend dry DNA with TE (10 mM Tris-HCl pH 8, 0.1 mM EDTA). Pro-Tip DNA resuspension can take...culture of bacteria transformed with your plasmid Resuspension buffer Denaturing solution Renaturing solution...several tubes/bottles. Remove the supernatant and resuspend the bacteria in buffer. Note: This step gets all... all of the bacteria back into suspension, but within a smaller volume of buffer that is compatible with...next solution. Add a denaturing solution to the resuspended bacteria. Note: This step causes the bacteria...column with 70% ethanol to remove excess salt. Resuspend the DNA pellet, or elute the DNA off of the column...Prepare the following solutions: Solution I - Resuspension Buffer 25 mM Tris-HCl (pH 8) 50 mM glucose 10...

  2. AAV Production in HEK293 Cells

    Type
    Protocol
    ...Pipet back and forth to resuspend each pellet completely. Combine resuspended pellets and keep on ice....Pipet back and forth to resuspend each pellet completely. Combine all resuspended cell pellets and sonicate...containing the resuspended virus from step 12 above. Add 50 units of benzonase per mL of viral suspension (you ...cells after 30 passages. Procedure Trypsinize and resuspend the HEK293T cells from 2 x T-175 flasks. Cells...vigorously multiple times to obtain a single cell suspension (no clumps of cells). Pool cells from 2 x T-175... Return to incubator for 48 h. Trypsinize and resuspend cells from the CS2. Cells should be at ~80% confluence...forth or shake vigorously to obtain a single cell suspension (no clumps). Count cells using a hemocytometer...

  3. Kit Free RNA Extraction

    Type
    Protocol
    ...tissues, extraction of RNA, precipitation, and resuspension. This protocol provides two options: using Solution...RNA in the bottom of the tube. Wash the RNA by resuspending the pellet in 0.5–1 ml of 75% Ethanol and vortex...important to not let the pellet get too dry before resuspending, as this affects the solubility of the RNA. ... but while the white pellet is still visible. Resuspend RNA pellet in RNase-free water or TE. Quantify...

  4. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...microcentrifuge tube. Vortex the cell suspension. Transfer 10 µL of cell suspension into the microfuge tube containing...culture. This protocol describes how to transfect suspension HEK293 cells with recombinant antibody plasmids... 5 mL pipette, transfer 0.5 mL of HEK293 cell suspension into a clean microcentrifuge tube. Pro-Tip Cells...Cells settle quickly and need to be resuspended before sampling. Gently swirl the flask 5–10 times before...Pipette 10 times to mix. Load 10 µL of the cell suspension/trypan blue mix into a cell counting chamber....

  5. Weighing Reagents Protocol

    Type
    Protocol
    ...of reagent that will be resuspended in a small volume (e.g. 0.02 g to resuspend in 1 mL), you may place...

  6. Western Blot

    Type
    Protocol
    ...5 min at 100 x g . Gently remove supernatant. Resuspend cell pellet in 1 mL of 1X PBS and transfer to ...min at 100 x g . Carefully remove supernatant. Resuspend cell pellet in appropriate volume of cold lysis...

  7. Lentivirus ddPCR Titration

    Type
    Protocol
    ... media and 11.1 µg/mL Polybrene. Mix the cell suspension well before seeding. Mix each well with a 1 mL...by incubating with 200 µL TrypLE for 1–2 min. Resuspend cells in 500 µL DMEM complete and transfer to ...

  8. DNA Quantification

    Type
    Protocol
    ...spectrophotometer using the solution the DNA is resuspended in, but with no DNA added. 'Blanking' measures...

  11. Fluorescence Titering Assay

    Type
    Protocol
    ... pipetting or inverting. Aliquot 2 mL of cell suspension into each well of the 6-well dish. Incubate the...

  13. Colony Formation Titering Assay

    Type
    Protocol
    ...pipetting or inverting. Aliquot 1.35 mL of the cell suspension into each well of the 6-well dish directly on...

  16. CRISPR Library Amplification

    Type
    Protocol
    ...cells are subject to lysis by freeze-thaw if not suspended in cryoprotectant like Glycerol or DMSO solutions...
Showing: 1 - 17 of 17 results