Skip to main content
Addgene

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

In Category

We narrowed to 286 results for: lum

Showing: 261 - 280 of 286 results

  1. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ...recommended by manufacturer) dH 2 O up to total volume Pro-Tips The amount of DNA that you cut depends... often requires 1 µg of DNA. The total reaction volume usually varies from 10-50 µL depending on application...application and is largely determined by the volume of DNA to be cut. A typical restriction digestion reaction...BSA (if recommended) x µL dH 2 O (to bring total volume to 30µL) The amount of restriction enzyme you use... enzyme because it is difficult to pipette less volume than this; 0.2-0.5 µL will likely be more enzyme...

  2. General Transfection

    Type
    Protocol
    ... 2 flask in a volume of 15 mL. Wednesday: Plate 1x10 6 cells in a T75 flask in a volume of 15 mL. Friday...Friday: Plate 8x10 5 cells in a T75 flask in a volume of 15 mL. Do not add antibiotics to the media. Use...depending on the user, quantities of chemical used, volumes, pH adjustment etc. Consequently, each batch needs...ratios to test: Ratio of DNA:PEI Amount of DNA (μg) Volume of 1 mg/mL PEI (μL) 1:1 18.9 18.9 1:2 18.9 37.8...

  3. Colony Formation Titering Assay

    Type
    Protocol
    .../mL polybrene: Dilution Volume of Lentivirus or Previous Dilution (μL) Volume of DMEM Complete Containing...Containing 10 μg/mL Polybrene (μL) Volume of Virus Added to Plate (μL) Volume of Cells Added to Plate (μL) ...mL) as follows: TU/mL = number of colonies/total volume in the well (mL) x dilution factor e.g., If the...

  4. AAV ddPCR Titration

    Type
    Protocol
    ...enough master mix for 9 samples. ITR Master Mix Volume 9X Volume Final Concentration 2X ddPCR Supermix for .../mL R = Reaction volume (20 µL) C = Copies/µL D = Dilution factor of virus V = Volume of virus in reaction...900 nM Nuclease-free water 5.9 µL 71.55 µL Total Volume 20 µL Vortex the master mix for 15 sec and spin...

  5. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...master mix,” which minimizes the number of small volumes you need to pipet. If you are using the same template...PCR product. Master Mix Preparation Multiply the volume of each reagent by the number of individual PCRs...reactions (7 unique primer pairs), so we multiply each volume by 7. In a single 1.5mL tube combine the following... 2.5µl (fwd primer) – 2.5µl (rev primer) = 45µl volume of master mix to add to each PCR tube. Secure the...

  6. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    .... Mix the dilutions well. Dilution Volume of Lentivirus (μL) Volume of DMEM complete + 10 µg/mL polybrene...cells as follows: Dilute 350,000 cells into a total volume of 7 mL of DMEM complete + 10 µg/mL polybrene. ... well of the 6-well dish. This brings the total volume in each well up to 1.5 mL. Since all the media ...

  7. Kit Free RNA Extraction

    Type
    Protocol
    ...disturbing the interphase layer, consider using a lower volume pipettor like a p200 to collect a majority of the...Chloride for this step. Isopropanol (Option A): Add 1 volume of Isopropanol to the extracted aqueous layer. ...

  8. CRISPR Library Amplification

    Type
    Protocol
    ... the Maxiprep protocol, scale the reagent volume and column number as needed. The use of Mega or Gigapreps... plateau and sometimes fall off entirely if the column or reagents are significantly overloaded. If you...Consult the manufacturers handbooks for appropriate volumes and numbers of tandem purifications. Visit our ...

  9. Protocol - How to Purify DNA from an Agarose Gel

    Type
    Protocol
    ...many kits cannot handle more than a certain total volume of gel per reaction. Place the gel in a labeled... the gel is directly proportional to its liquid volume and this is used to determine how much of each ...sample you want to load on a gel, make 10% more volume than needed because several microliters can be ...

  10. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...1 1 μL AgeI add ddH 2 O to bring to 50 μL final volume Incubate at 37°C for 2 hours. Purify with Qiaquick... DNA ligase add ddH 2 O to bring to 20 μL final volume Incubate at 16°C for 4-20 hours. Transform 2 μL...0.8 μL NcoI add ddH 2 O to bring to 20 μL final volume Incubate at 37°C for 1-2 hours. 4. Run the digestion...master mix to each tube from step c for a total volume of 100 μL. Pipette master mix directly into the...

  11. Pouring LB Agar Plates

    Type
    Protocol
    ...200 mL of agar in 500 mL bottles. The extra empty volume is necessary to prevent your molten agar from boiling...Cover the opening of the bottle with its cap or aluminum foil (but do not make an air-tight seal!) and ...the first plate to get a good idea of what that volume looks like in your particular plate. For the remainder...

  12. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...loosely cover the entire top of the bottle with aluminum foil. Autoclave and allow to cool to room temperature... swirl. Loosely cover the culture with sterile aluminum foil or a cap that is not air tight. Incubate ...

  13. Ligation Independent Cloning

    Type
    Protocol
    ...Inactivate T4 Pol by heating to 75° for 20 minutes. Volume (μl) Reagent Final Conc. 4 10X Buffer 1X 20-30 ...annealing reaction. This should be done in a small volume with no additional water (<5 μl). It is advisable...

  14. Centrifugation

    Type
    Protocol
    ...often an extra tube filled with water to the same volume as your samples. If your samples are significantly...need to fill the blank based on weight instead of volume. Pro-Tip For microfuge tubes, place them with the...

  16. Protocol - Bacterial Transformation

    Type
    Protocol
    ...you to recover all transformants. If the culture volume is too big, gently collect the cells by centrifugation...centrifugation and resuspend in a smaller volume of LB so that there isn't too much liquid media on the agar...

  17. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...of HCl at a time. Add deionized water to a final volume of 1 L and recheck pH to ensure that it has not...up to 4 times for a total of 16% of the culture volume. At 72-96 h post-transfection, supplement the flask...

  18. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ... . Simply adjust the mass of agarose in a given volume to make gels of other agarose concentrations (e.g...sample you want to load on a gel, make 10% more volume than needed because several microliters can be ...

  19. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...to a microfuge tube. Add 8 µL of PBS to a total volume of 13 µL . Prepare the recombinant antibody Dilute... issues. Plot the mg/mL versus the HC + LC Area column for the IgG standards in Google Sheets. Open exported...

  20. DNA Quantification

    Type
    Protocol
    ...purity. Note: Purity is measured under the 260/280 column (A good purity ranges from 1.80-2.00). Repeat for...
Showing: 261 - 280 of 286 results