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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 37 results for: UTY

Showing: 1 - 10 of 37 results

  1. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ... a 1:100 dilution of the homogenized cell solution by adding 10 µL homogenized cell solution to 990 µL...Cloning by Limiting Dilution Isolating a Monoclonal Cell Population by Limiting Dilution You may also like...cell solution later in this protocol. Isolating a Monoclonal Cell Population by Limiting Dilution Isolate...homogenized cell solution is transferred. Transfer cells from the homogenized cell solution into the conditioned...cell solution at a concentration of 5 cells/mL. Prepare approximately 10 mL of this cell solution for ...medium, prepare this cell solution in fresh complete medium. This 5 cells/mL solution will be used to seed ... quantitate the homogenized cell solution. Homogenized cell solution concentration: 4 × 10 5 cells/mL ...

  2. AAV ddPCR Titration

    Type
    Protocol
    ...in the dilution BSC. Prepare 1X dilution buffer (see recipe above). Save ~25 µL of 1X dilution buffer ... carefully add the dilution buffer to the dilution plate according to the dilution scheme listed in step...Pro-Tip Prepare the viral dilutions in a separate biological safety cabinet (dilution BSC) than that used for...touching the screen. Prepare the Serial Dilution Place a 48-well dilution plate in a chilled 96-well freezer... add 5 µL of each viral sample to Dilution 1 in the 48-well dilution plate and pipette 5–10 times to mix...change tips each time. Dilution 1 (20X): 5 µL in 95 µL 1X PCR buffer (1:20) Dilution 2 (20X): 5 µL in 95 ...PCR buffer (1:400) Dilution 3 (20X): 5 µL in 95 µL 1X PCR buffer (1:8,000) Dilution 4 (20X): 5 µL in 95...

  3. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...of 2 x 10 8 dilution 90 2 x 10 7 10 of 2 x 10 7 dilution 90 2 x 10 6 10 of 2 x 10 6 dilution 90 2 x 10 ...Total dilution Dilution 1 (DNase step) 5 uL AAV stock 45 uL 10X 10X Dilution 2 5 uL Dil. 1 95 uL 20X 200X...200X Dilution 3 20 uL Dil. 2 80 uL 5X 1000X Dilution 4 20 uL Dil. 3 80 uL 5X 5000X Dilution 5 20 uL Dil...25000X Dilution 6 20 uL Dil. 5 80 uL 5X 125000X Dilution 7 20 uL Dil. 6 80 uL 5X 625000X Dilution 8 20 ... To obtain a solution at 2 x 10 9 molecules/μL: 1.59 x 10 11 / 2 x 10 9 = 79.8X dilution ...10 5 10 of 2 x 10 5 dilution 90 2 x 10 4 10 of 2 x 10 4 dilution 90 2 x 10 3 Pro-Tip To help stabilize ... standard dilution. Dilute DNase-treated and AAV reference samples according to the dilution scheme in...

  4. Lentivirus ddPCR Titration

    Type
    Protocol
    ...transfer 200 µL of the first dilution into the next dilution. Repeat for all dilutions as indicated in the chart... virus dilutions were used to transduce cells: Initial dilution factor of virus stock Dilution factor ... µL 5 200 µL of 2.5-fold dilution 200 µL 400 µL 10 200 µL of 5-fold dilution 200 µL 400 µL 20 200 µL of... of 10-fold dilution 200 µL 400 µL 40 200 µL of 20-fold dilution 200 µL 400 µL 80 200 µL of 40-fold dilution... 160 200 µL of 80-fold dilution 200 µL 400 µL Add 150 µL of each viral dilution to a well of a 6-well ... viral dilution). Remember, this is an additional 10-fold dilution of the virus stock! Don’t forget to...~2 as you progress from the higher dilutions to the lower dilutions (blue). As shown in the image (Figure...

  5. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...EDTA Store Solution I at 4°C Solution II - Denaturing Solution 0.2 N NaOH 1.0% SDS Store Solution II at room...plasmid Resuspension buffer Denaturing solution Renaturing solution 2 mg/mL RNase A TE or water-saturated...that is compatible with the next solution. Add a denaturing solution to the resuspended bacteria. Note...contents, including plasmid DNA, into solution. Add a renaturing solution to the denatured bacteria. Note: ...Lysis Plasmid Miniprep Prepare the following solutions: Solution I - Resuspension Buffer 25 mM Tris-HCl (pH... room temperature Solution III - Renaturing Solution (Potassium Acetate) 120 mL 5M Potassium acetate 23...Resuspend the pellet in 100 μL of cold Solution I. Vortex the solution for 2 min or until all bacteria are...

  6. Colony Formation Titering Assay

    Type
    Protocol
    ... Mix the dilutions well Note: the 1:10 dilution can usually be omitted because this dilution typically... serial dilutions of the lentivirus into DMEM complete containing 10 μg/mL polybrene: Dilution Volume ...quantified. Add 150 μL of a viral dilution to each well (each well gets one dilution) Also include an untransduced... colonies for at least 2 of the dilutions. Pro-Tip The higher dilution wells tend to provide more accurate...present in the lower dilutions, and single colonies are visible in the higher dilutions. Last reviewed on...staining resistant colonies with crystal violet solution and, therefore, the cells cannot be used for later...DMEM complete containing 10 μg/mL polybrene by diluting 20 μL of 10 mg/mL polybrene into 20 mL of media...

  7. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...different incubation options available. Caution Use the Stop Solution in a well ventilated area and avoid ...Immediately before use, mix 5 mL TMB Solution and 5 mL Peroxide Solution from the TMB substrate kit. Cap the...determine the ideal concentration. Prepare serial dilutions of the purified antigen as follows: 2 ng/µL : ... wells of each row, using a different standard solution for each row of a 96-well ELISA microplate. Cover...the wash buffer from the wells. Prepare antibody dilution buffer (0.1% BSA in PBS) as follows: Dilute 50... to the desired concentrations in the antibody dilution buffer. Pro-Tip The ideal antibody concentration...multichannel pipette, add 100 µL of primary antibody solution to the appropriate wells. Cover the plate with...

  8. Kit Free RNA Extraction

    Type
    Protocol
    ...Option #1 - Solution D Protocol Before starting this protocol, make sure to prepare solution D (see reagent...or lyse tissues or cells in Solution D. For tissues: use 1 mL of Solution D per 100 mg of cells. For cultured...resuspension. This protocol provides two options: using Solution D (see reagent section for recipe) or using an...all-in-one acid guanidinium thiocyanate-phenol solution such as TRIzol®. For the precipitation step, two...reagents RNase-free (Use an RNase decontamination solution, such as RNaseZap® or RNase AWAY®, may be used... Vortexer -20°C freezer -80°C freezer Reagents Solution D (for Protocol Option #1 ): 4 M Guanidinium Thiocyanate...alcohol (49:1) 75% Ethanol RNase-free water or TE solution RNase free tubes: microcentrifuge tubes, 4 mL ...

  9. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...viral dilution to each well (each well gets one dilution, so a 6-well plate will hold 5 dilutions plus ...just a sample of possible dilutions. You may want to try higher/lower dilutions depending on your downstream...the results of your titration. Mix the dilutions well. Dilution Volume of Lentivirus (μL) Volume of DMEM...that already contain 0.5 mL of virus solutions at various dilution. Make sure to use the polybrene-containing...batch of DMEM complete + 10 µg/mL polybrene by diluting 20 µL of 10 mg/mL polybrene into 20 mL media. ...aliquot on ice prior to use. Prepare a range of dilutions of the lentivirus in DMEM complete + 10 µg/mL ...polybrene-containing media to make the cell solution in this step. To seed the cells: Prepare a batch of cells as ...

  10. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...swirling the flask occasionally as the solution heats up.). Caution HOT! Be careful stirring, eruptive boiling...do you get better resolution of bands? A few simple ways to increase the resolution (crispness) of your...gel electrophoresis, including tips to improve resolution and separation of bands. Protocols... completely dissolved (but do not overboil the solution, as some of the buffer will evaporate and thus...continue towards a boil. Keep an eye on it the solution has a tendancy to boil over. Placing saran wrap...necessary if you pay close attention. Let agarose solution cool down to about 50 °C (about when you can comfortably...0.2-0.5 μg/mL (usually about 2-3 μl of lab stock solution per 100 mL gel). EtBr binds to the DNA and allows...
Showing: 1 - 10 of 37 results