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We narrowed to 1,076 results for: TOR

Showing: 801 - 820 of 1076 results

  1. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...much virus as possible. Store at 4 °C for short term (2 weeks), or aliquot and store at -80 °C for long term... Ultracentrifugation You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Introduction... one (long) day. Equipment Ultracentrifuge T70i rotor or equivalent Reagents Opti-Prep (60% Iodixanol)...Sterilize by passing through a 0.22 μm filter and store at 4 °C. 1X PBS-MK buffer Dissolve 26.3 mg of MgCl...Sterilize by passing through a 0.22 μm filter and store at 4 °C. 0.1% Pluronic-F68 in PBS (A) Add 500 µL... Centrifuge at 350,000 x g for 90 min in a T70i rotor at 10 °C. Pro-Tip If you need more time, you can.... Carefully take the QuickSeal tubes out of the rotor and place them in a stable rack. **Make sure not...

  2. AAV ddPCR Titration

    Type
    Protocol
    ...Protocols ddPCR Titration of AAV Vectors ddPCR Titration of AAV Vectors You may also like... AAV Titration...Titration by qPCR AAV Viral Service Viral Vector Guides Virus Blog Posts Molecular Biology Reference Introduction...titer purified recombinant Adeno-associated viral vectors (AAV). This protocol specifically uses primers ...probes targeting the ITR elements in the viral vectors but can be modified for other targets. The dilution... Transfer the cartridge holder to the droplet generator. Close the lid and wait for the droplets to be...Reaction volume (20 µL) C = Copies/µL D = Dilution factor of virus V = Volume of virus in reaction mix (4...concentration obtained by ddPCR should decrease by a factor of 2 across the dilutions. In the example below...

  3. Centrifugation

    Type
    Protocol
    ... the two, use a conversion calculator or chart like the G Force calculator (Link opens in a new window...Introduction A centrifuge is a common piece of laboratory equipment used to separate different components...tabletop microcentrifuge with the lid open, the rotor with microfuge tubes in place, and the control panel...centrifuge, with tubes evenly distributed around the rotor. How to Centrifuge Now that you know the different...working smoothly. Place your sample tubes into the rotor so that they are distributed evenly around the center...sample. Place the centrifuge’s internal lid on the rotor (if it has one) and then close the external lid....

  4. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...polymerase pLKO.1 - TRC Cloning Vector Cloning protocols for using the pLKO.1 vector, a backbone used by the ...addgene.org with any questions. Intro to the Lab Bench Introductory techniques designed to help you get started ...and Two (BSL-1 and BSL-2) Safety measures for laboratories operating at BSL-1 and BSL-2 Watch the Video...Video! Pipetting Learn about selecting the correct pipettor and pipette tip, how to dispense liquids, and ... its use Weighing Reagents Learn how to weigh laboratory materials on a balance Basic Molecular Biology...Watch the Video! Creating Bacterial Glycerol Stocks Store bacterial strains or plasmids for long term use ...

  5. Fluorescence Titering Assay

    Type
    Protocol
    ...Assay for Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...marker. Determining the titer of your lentiviral vector allows you to control the multiplicity of infection...1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Inverted...heat-inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS...protocol was developed using low titer lentiviral vectors (10 5 TU/mL). If you anticipate that your viral...units per mL (TU/mL) using either the dilution factors (method 1) or the volume of virus (method 2): Method...Fraction of cells with fluorescence D = Dilution Factor V T = Transduction Volume, mL For example: If 150,000...

  6. General Transfection

    Type
    Protocol
    ...filter. Aliquot 50–100 µL and store at -20 °C. Aliquots can be thawed and stored at 4 °C prior to use. Thawed...into sterile tubes. Store the tubes at -80 °C. After thawing the solution can be stored at 4 °C for up to...General Transfection You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents...heat-inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS...

  7. Colony Formation Titering Assay

    Type
    Protocol
    ...Assay for Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents...heat inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS...Swirl gently to mix and place the dish in the incubator. Incubate for 48–72 h. Pro-Tip Different cells...colonies/total volume in the well (mL) x dilution factor e.g., If the 1:100,000 well has 75 colonies, then...the indicated serial dilutions of the lentiviral vector pRosetta . The following day, cells were treated...the indicated serial dilutions of the lentiviral vector pHAGE EF1α dCas9-KRAB . The following day, cells...

  9. Pipetting Protocol

    Type
    Protocol
    ...volume. Below that is the tip ejector button which is used to push the tip ejector down the pipette to remove...Learn about selecting the correct pipettor and pipette tip, how to dispense liquids, and how to handle...liquid (think: 0.1 µL to 1 mL). When working in a laboratory, properly dispensing liquid ensures the accuracy...the set volume for the the pipette, and the tip ejector ring at the bottom which pushes the pipette tip...dedicated waste container and pressing on the tip ejector button. If pipetting biohazardous materials, be...

  11. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ... plates) and store at -20 °C. Once a standard is thawed do not freeze it again but store at 4 °C and use...SYBR Green Technology You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Introduction...the titer of samples obtained from academic viral vector cores. Our titers were similar to those reported...30 min Equipment qPCR instrument Heating plate Pipettors 1–10 µL single channel pipette 20–200 µL single...uL) Volume of nuclease free water (uL) Dilution factor Total dilution Dilution 1 (DNase step) 5 uL AAV...

  13. The 12 Days of CRISPR

    Type
    Blog Post
    ...CRISPR, Addgene gave to me: An engineered prime editor with enhanced editing efficiency in plants 🎵 ...

  15. New Videos: Addgene Lab Tips

    Type
    Blog Post
    ...Addgene Videos Page 5 Science Rap Videos and Their Creator: Tom McFadden Using Video to Share Your Science...

  20. CRISPR Library Amplification

    Type
    Protocol
    ...amplification protocols that have been developed by the depositor for a specific pooled library available at Addgene...Addgene. We recommend following the depositor's specific protocol if one is available. If a pooled library...require modifications dictated by the originating laboratory for optimal results. If you obtained the pooled...amplification protocol is expected to obtain satisfactory results for libraries up to 200,000 individual...limited. Equipment Table top centrifuge BioRad Electroporator (MicroPulser TM , Bio-Rad 1652100) Reagents...time for a total of eight electroporations): Electroporator Conditions: Bio-Rad Micropulser Ec1 0.1 cm ...
Showing: 801 - 820 of 1076 results