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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 728 results for: STO;

Showing: 701 - 720 of 728 results

  1. AAV Production in HEK293 Cells

    Type
    Protocol
    ...into sterile tubes. Store the tubes at -80 °C. After thawing, the solution can be stored at 4 °C for up to... glutaGRO 11 mL of 200 mM L-alanyl-L-glutamine. Store at 4 °C. D1 + 0.1 M sorbitol (optional) : DMEM ...months, discard the tube and thaw a new working stock. 40% POLYETHYLENE GLYCOL (PEG) 8000 solution: Dissolve... solution may separate into phases. Aliquot and store at 4 °C. Cell Lysis Buffer : 50 mM Tris HCl, 150....5 Filter sterilize through a 0.22 μm membrane. Store at 4 °C. Considerations Before You Start The health...

  2. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...Agar Plate Creating Bacterial Glycerol Stocks for Long-term Storage of Plasmids Recovering Plasmid DNA from... For long term storage of the bacteria, you can proceed with Creating a Glycerol Stock . You can now isolate...temperature. Now screw on the top of the bottle and store the LB at room temperature. When ready to grow your...

  3. Lentivirus ddPCR Titration

    Type
    Protocol
    ...Prepare 25 ng/µL stocks of each sample. Samples can be used for ddPCR immediately or stored at -20 °C until...For the first virus dilution, add 160 µL of viral stock to 240 µL of media. When adding the virus to the...Dilute Media Volume Total Volume 2.5 160 µL viral stock 240 µL 400 µL 5 200 µL of 2.5-fold dilution 200 ... is an additional 10-fold dilution of the virus stock! Don’t forget to factor this into your calculations...transduce cells: Initial dilution factor of virus stock Dilution factor after additional 10-fold dilution...

  4. Educational Resources

    Type
    Guide
    ...Transformation Plasmid Cloning Creating Bacterial Glycerol Stocks Restriction Digests Guides Browse our in-depth ...

  5. 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...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% Poloxamer 188 in PBS (A) Add 500 µL...clean fractions. Concentration and buffer exchange: Stocks of Poloxamer 188 Poloxamer 188, 10% solution A:...

  6. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ... mM EDTA Store Solution I at 4°C Solution II - Denaturing Solution 0.2 N NaOH 1.0% SDS Store Solution ...acetate 23 mL glacial acetic acid 57 mL of dH 2 O Store Solution III at 4°C Grow 2 mL overnight cultures...containing your plasmid of interest. Add 1.5 mL of the stock culture to a 1.75 mL microfuge tube. Centrifuge ... room temperature before quantifying and using. Store DNA at 4°C. Tips and FAQ Plasmid purification kits...

  7. Protocol - How to Streak a Plate

    Type
    Protocol
    ...Bacterial Culture Introduction If you have a glycerol stock or stab culture of bacteria and need to purify plasmid...population (single colony) of bacteria from this stock. Using a single colony from a freshly streaked agar...Obtain the approrpriate bacterial stab or glycerol stock . Using a sterile loop, pipette tip or toothpick... of the stab culture or the top of the glycerol stock. Pro-Tip If you use a wire loop you can sterilize...

  8. Fluorescence Titering Assay

    Type
    Protocol
    ...viral stock will be higher titer, consider additional dilutions. Dilution Volume of Lentivirus Stock (μL...heat-inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS...it is recommended that you titer from the frozen stock to account for any loss in titer associated with...

  9. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...Reagents TAE ( recipe here ) Agarose Ethidum bromide (stock concentration of 10 mg/mL) Gel loading buffer, New...Pro-Tip It is a good idea to microwave for 30-45 sec, stop and swirl, and then continue towards a boil. Keep...approximately 0.2-0.5 μg/mL (usually about 2-3 μl of lab stock solution per 100 mL gel). EtBr binds to the DNA ...sample is unloaded, push the pipettor to the second stop and carefully raise the pipette straight out of ...

  10. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ... The above graph displays the stock concentration of Carbenicillin stock used (150 µL per plate). Please... concentration (100 mg/mL, 1000x) carbenicillin stock solution in sterile water (or other antibiotic) ...

  11. Optogenetics Guide

    Type
    Guide
    ...cells. LITE Two-hybrid system integrating the customizable TALE DNA-binding domain with cryptochrome 2 ...

  12. Protocol - Bacterial Transformation

    Type
    Protocol
    ...because bacteria are used as the means for both storing and replicating plasmids. Because of this, nearly...for transforming plasmid DNA for the purposes of storage and amplification. Higher efficiency cells are ... (containing the appropriate antibiotic ) from storage at 4°C and let warm up to room temperature and ...

  13. Colony Formation Titering Assay

    Type
    Protocol
    ...heat inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS...it is recommended that you titer from the frozen stock to account for any loss in titer associated with... to Plate (μL) Final Viral Dilution 1:10 100 of Stock Virus 900 150 1,350 1:100 1:100 100 of 1:10 900 ...

  14. Protocol - How to Perform Sequence Analysis

    Type
    Protocol
    ...posted sequence in the "View Sequence" link. Is your custom primer found within the Addgene sequencing results...require additional sequencing and need to design a custom primer, Addgene recommends using Addgene's sequencing...

  15. Ligation Independent Cloning

    Type
    Protocol
    ... reaction, in which a single dNTP is included to stop the exonuclease function of the enzyme and shift...vector (shown in blue here). The T4 reaction may be stopped by heating to 75° for 20 minutes. Step 6: Anneal...

  16. Video Library

    Type
    Protocol
    ...enzymes Diagnostic Restriction Digest Protocol Storing and Handling Addgene Plasmids What to do after ...receive your plasmids from Addgene Instructions for Storing and Handling Plasmids Genomic Deletions with CRISPR...

  18. Immunocytochemistry

    Type
    Protocol
    ...nM DAPI working solution: Prepare a 300 µM DAPI stock solution by diluting 2.1 µL of the 5 mg/mL DAPI ...working solution by diluting 5 µL of the 300 µM DAPI stock solution into 5 mL PBS. Protect from light. Procedure...

  19. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ... heat-inactivated FBS and 5mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and FBS lots can...and have been sufficiently expanded, prepare cell stocks and/or harvest to test for protein expression. ...

  20. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ... want to clone from the start codon (ATG) to the stop codon (TGA, in this example). Assuming you are ...taking the final 18bases of the ORF, including the stop codon (5'-TGGCATATCTCGAAGTACTGA-3'), then adding...
Showing: 701 - 720 of 728 results