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

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We narrowed to 712 results for: STO

Showing: 681 - 700 of 712 results

  1. 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...standard curve plasmid (2 x 10 9 stock made in step #1): Volume of 2 x 10 9 stock or previous dilution (μL) ...Water 4.7 μL 470 μL Procedure Prepare a plasmid stock of 2 x 10 9 molecules/μL to generate a standard ...79.8 = 1.25 μL Therefore we need to dilute 1.25 μL stock into 98.74 μL H 2 0 Pro-Tip Once a validated standard...Total dilution Dilution 1 (DNase step) 5 uL AAV stock 45 uL 10X 10X Dilution 2 5 uL Dil. 1 95 uL 20X 200X...

  2. Pouring LB Agar Plates

    Type
    Protocol
    ...your plates are ready for use or storage at 4 ℃. You should not store your plates for longer than 1 month...Antibiotic Concentrations Antibiotic Recommended Stock Concentration Recommended Working Concentration ...to the molten gel mix, you should create a 1000x stock solution. For example: If you’ll be preparing plates...concentration of 100 ug/mL ampicillin, you should make a stock solution of 100,000 ug/mL (100 mg/mL). Simply measure...solidifies in the bottle while you’re pouring, you should stop pouring and re-make the gel-mix. If you’re making...material to reduce condensation. The plates are then stored at 4 ℃ until use. Once your plates have solidified...

  3. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...to a polypropylene storage tube. The media contains your lentiviral particles. Store at 4°C. m. Add 5 mL...may be stored at 4°C for a few days, but should be frozen at -20°C or -80°C for long-term storage. TIP:...concentration of 100 μg/mL antibiotic. Pour plates and store at 4°C. Hexadimethrine Bromide (Polybrene) Prepare...catalog #H9268) in 0.9% NaCl. Autoclave to sterilize. Stock solution is stable at 4°C for up to one year. The...stable at 4°C for several years. Protamine Sulfate Store protamine sulfate (MP Biomedicals catalog #194729...soluble in cold water. Puromycin Prepare a 50mg/mL stock solution of puromycin (Sigma-Aldrich catalog #P8833... Sterilize by passing through a 0.22 μm filter. Store aliquots at -20°C. I.3. Warranty Information Addgene...

  4. 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...heat-inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS...months, discard the tube and thaw a new working stock. Considerations Before You Start The health of the...

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

    Type
    Protocol
    ... sterile sodium phosphate dibasic (NaH 2 PO 4 ) Store up to 1 month at 4 °C Adjust pH to 7.0 Autoclave...Gravitrap columns and gently pour off the ethanol storage buffer. Tightly attach LabMate PD10 Buffer Reservoirs...washing with 25 mL of Protein A/G binding buffer and store in 20% ethanol at 4 °C . Pro-Tip Columns may be ...Centrifuge column at 1000 x g for 2 min to remove storage solution. Place a mark on the side of the column...antibody to 1 mg/mL with PBS if needed. For long term storage, add sterile sodium azide to 1 mM. Option 2: Buffer... the NanoDrop Spectrophotometer. For long term storage, add sterile sodium azide to 1 mM....

  6. Kit Free RNA Extraction

    Type
    Protocol
    ...equipment back to room temperature, as prolonged storage in the cold room may damage it. Using a pipettor...quantification , which can be modified for RNA. Store your RNA sample(s) at -80 °C to prevent RNA degradation...sample, consider making smaller aliquots of it and storing those in -80°C. Option #2 - TRIzol® Protocol Homogenize...DNA quantification , which can be modified for RNA.Store your RNA sample(s) at -80 °C to prevent RNA degradation...sample, consider making smaller aliquots of it and storing those in -80°C. Solution D Protocol TRIzol® Protocol...

  7. 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...

  8. 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...

  9. 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...

  10. 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% Pluronic-F68 in PBS (A) Add 500 µL...clean fractions. Concentration and buffer exchange: Stocks of Pluronic F68 Pluronic F68, 10% solution A: 0.1%...

  11. 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...

  12. 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...

  13. 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...

  14. 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 ...

  15. 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) ...

  16. 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 ...

  17. 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 ...

  18. 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...

  19. 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...

  20. 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...
Showing: 681 - 700 of 712 results