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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 457 results for: nsf

Showing: 421 - 440 of 457 results

  1. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...recipient plasmid backbone. Transformation Proceed with the transformation according to the manufacturer...from your transformation will give you the first indication as to whether your transformation worked. Your...Digest of Plasmid DNA DNA Ligation Bacterial Transformation Summary PCR based cloning is incredibly versatile...competent cells. For most standard cloning, you can transform 1-2μl of your ligation reaction into competent...conduct a positive control to ensure that your transformation worked. You could also try varying the amount...

  2. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ... volume of homogenized cell solution is transferred. Transfer cells from the homogenized cell solution...990 µL complete medium. Instead of transferring 0.125 µL, now transfer 100 times that volume, which is 12.5... solution, transfer 12.5 µL of the 1:100 dilution to 10 mL of conditioned medium. Transfer 100 µL of the...before counting the cells. When you ultimately transfer approximately 50–100 cells to the conditioned ...

  3. Ligation Independent Cloning

    Type
    Protocol
    ...now ready for transformation. Use 1-2 μl of annealing reaction for each transformation, following a standard...stability to hold the plasmid together through the transformation/replication process. LIC employs long overhangs...association between fragments, allowing for transformation without ligation. Because of its dual polymerase...heating to 75° for 20 minutes. Step 6: Anneal and Transform Mix your treated vector and insert at a molar ...

  4. Video Library

    Type
    Protocol
    ... tips Bacterial Transformations We walk you through a standard plasmid transformation protocol and offer...tricks to reduce transformation time and increase efficiency Bacterial Transformation Protocol Inoculating...when requesting samples from Addgene Technology Transfer How to Order A step-by-step walkthrough of Addgene's...

  5. Lentivirus ddPCR Titration

    Type
    Protocol
    ... pipette 10–20 times. Use a new pipette tip to transfer 200 µL of the first dilution into the next dilution...min. Resuspend cells in 500 µL DMEM complete and transfer to a microcentrifuge tube. Centrifuge for 5 min...the DG8 gasket, making sure that it is secure. Transfer the cartridge holder to the droplet generator....while slowly and gently pipetting the droplets. Transfer the droplets to a prechilled PCR plate. Pro-Tip... 98 10 2 1 Hold 4 ∞ 2 1 After PCR is complete, transfer the plate to the Droplet Reader. Open the QuantaSoft...

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

    Type
    Protocol
    ...Ligation Assemble DNA using DNA ligase Bacterial Transformation Introduce DNA into a bacterial strain Watch...preparations. Name Description Link to Video General Transfection Introduce plasmid DNA to mammalian cells Lentivirus...Produce lentivirus with a polyethyenimine (PEI) transfection protocol Fluorescence Titering Assay for Lentivirus...applications. Name Description Link to Video Transfection Introduce plasmid DNA to mammalian cells to ...

  7. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ...Restriction Digest of Plasmid DNA Bacterial Transformation Background Information The final step in the...the backbone and the complete plasmid can be transformed into bacterial cells for propagation. The majority... incubation at 37°C. Proceed with bacterial transformation . Tips and FAQ Do controls When doing ligations...contamination of intact plasmid in ligation or transformation reagents Optimizing the Vector:Insert Ratio...

  8. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ... protein expression observed using transient transfection approaches, generating cell lines using lentiviral... stable cell line, as opposed to transiently-transfected cells, increases reproducibility, as it eliminates...variation associated with repeated transient transfection. Some lentiviral vectors deliver mammalian antibiotic...Different brands and FBS lots can promote or inhibit transfection. Test a variety of brands and FBS lot numbers...

  9. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...Collection Antibody Blog Posts Antibody Guide Transfection for Recombinant Antibodies Recombinant Antibody....125 mg/mL: 10 µL of 0.25 mg/mL with 10 µL PBS Transfer 5 µL of each standard to a microfuge tube. Add... antibody to a concentration of 0.9–1.1 mg/mL. Transfer 5 µL of recombinant antibody to a microfuge tube...

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

    Type
    Protocol
    ...Collection Antibody Blog Posts Antibody Guide Transfection for Recombinant Antibodies Immunocytochemistry...recombinant antibody to room temperature (see Transfection for Recombinant Antibody protocol ). Pro-Tip... has reached the desired concentration. Gently transfer the sample from the reservoir to a LoBind tube...

  11. AAV ddPCR Titration

    Type
    Protocol
    ...the DG8 gasket, making sure that it is secure. Transfer the cartridge holder to the droplet generator....while slowly and gently aspirating the droplets. Transfer the droplets to a prechilled PCR plate. Pro-Tip... 98 10 2 1 Hold 4 ∞ 2 1 After PCR is complete, transfer the plate to the Droplet Reader. Open the QuantaSoft...

  12. Kit Free RNA Extraction

    Type
    Protocol
    ...cells. Extract RNA from the homogenized sample(s).Transfer tissue/cell lysate to a 4 mL tube. Add the following...the cold room may damage it. Using a pipettor, transfer the top, aqueous phase to a new RNAse-free tube...

  13. Protocol - How to Create a Bacterial Glycerol Stock

    Type
    Protocol
    ... of Plasmids You may also like... Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture... not need to obtain more competent cells and retransform. Bacteria on an LB agar plate can be stored at...

  15. Weighing Reagents Protocol

    Type
    Protocol
    ...cross-contamination with other substances. They also help you transfer the material to your tube or container. Pro-Tip...weighed out the correct amount of your reagent, transfer it into your container. Make sure that any spills...

  16. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...Bacterial Culture You may also like... Bacterial Transformation Agarose Gel Electrophoresis Agarose Gel DNA...optional) Reagents Overnight culture of bacteria transformed with your plasmid Resuspension buffer Denaturing...

  17. Pouring LB Agar Plates

    Type
    Protocol
    ...x (0.220 L) = 8.14 g pre-mixed LB-agar powder. Transfer the LB-agar powder you’ve measured out into an...molten agar from boiling over in the autoclave. Transfer the sterile water (in our case 220 mL) to the ...

  18. Protocol - How to Streak a Plate

    Type
    Protocol
    ...also like... Making LB Agar Plates Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture...

  19. Fluorescence Titering Assay

    Type
    Protocol
    ...brands and lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to...

  20. Gibson Assembly Protocol

    Type
    Protocol
    ...You can purchase master mix or make your own. Transform bacteria with the DNA and screen for the correct...
Showing: 421 - 440 of 457 results