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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: 401 - 420 of 457 results

  13. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...a donor in FRET (fluorescence resonance energy transfer) and FLIM (Fluorescence Lifetime Imaging Microscopy...

  14. Lentivirus Production

    Type
    Protocol
    ...lentiviral vector transfected into 293T cells using a polyethylenimine (PEI) transfection protocol. This ...packaging cells Day 1 (pm): Transfect packaging cells Day 2 (am): 18 h post-transfection. Remove media, replace...filter Transfection grade DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and Transfer plasmid... optimized for each transfer plasmid. Pro-Tip Endotoxins can inhibit transfection; therefore, plasmid ...modified for alternative packaging cell lines or transfection reagents. Once produced, lentivirus can be used...brands and lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to... 2 for ~20 h. Prepare a mixture of the three transfection plasmids into OptiPro SFM: Reagent Amount per...

  15. Western Blot

    Type
    Protocol
    ...protocol uses a dry transfer device but can be adapted for wet and semi-dry transfer approaches. Last Update...well. Assemble the transfer sandwich as follows: Unseal the iBlot 2 PVDF Mini transfer stack. Set the Top...iBlot 2 Gel Transfer Device. Wet the pre-run gel in deionized water and place it on the transfer membrane... parameters are correct. Transfer for 5–6 min for proteins <30 kDa. Transfer for 8–10 min for proteins...Timeline Day 1: Prepare lysates, run SDS-PAGE, transfer, block, incubate with primary antibody Day 2: ...Mini gel tank chamber Power supply iBlot 2 Gel Transfer Device Roller Spatula Platform shaker Cold room...supernatant. Resuspend cell pellet in 1 mL of 1X PBS and transfer to a microcentrifuge tube. Centrifuge for 5 min...

  16. AAV Production in HEK293 Cells

    Type
    Protocol
    ...cells in CS2 Day 2: Seed cells in CS5 Day 3 (am): Transfect cells Day 7 (am): Harvest cells Equipment Class...brands and lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to...Polyethylenimine (PEI) 25 kDa MW Pro-Tip Other transfection reagents may be used in this protocol, but their...their conditions must be optimized. Plasmids for transfection: pHelper; pRC (Rep-Cap), plasmid expressing ... Pass and plate the cells the day before the transfection. Thaw a new vial of cells after 30 passages....cells, add 200 mL of DMEM complete media, then transfer the cells into a sterile bottle. Rinse the CS2...to the incubator for 24 h–36 h. Proceed with transfection: Calculate the amount of each plasmid needed...

  17. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...Day 2: b. Perform the transfection in the late afternoon because the transfection mix should only be incubated...Digesting pLKO.1 TRC-Cloning Vector C.4 Ligating and transforming into bacteria D. Screening for Inserts D.1 Recommended...pLKO.1 can be introduced into cells via direct transfection, or can be converted into lentiviral particles...Measure the DNA concentration. C.4 Ligating and Transforming into Bacteria Use your ligation method of choice...final volume Incubate at 16°C for 4-20 hours. Transform 2 μL of ligation mix into 25 μL competent DH5 ...transient knockdown of protein expression, you may transfect plasmid DNA directly into the target cells. The...#12259 HEK-293T cells GenHunter: #Q401 FuGENE® 6 Transfection Reagent Roche Applied Biosciences: #11814443001...

  18. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ...incubation, transform DH5α E. coli cells by heatshock with the plasmid of interest. See our transformation page...for a detailed heatshock transformation protocol. Plate 50 µL of transformed E. coli /rescue media suspension...also like... Making LB Agar Plates Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture...plating antibiotic over-agar for the selection of transformed E. coli . Over-agar spreading of antibiotic makes...investigator to conveniently plate and select transformed cells containing plasmids differing in their ... sterile water (or other antibiotic) E. coli transformed with a plasmid containing the carbenicillin (...carbenicillin, plated over-agar for selection of transformed E. coli . Sample Data Selection of E.coli on ...

  19. CRISPR Library Amplification

    Type
    Protocol
    ... Day 1: Transform, recover, set up overnight growth (Estimated time 2-3 hours) Transformation should be...of 5 mL (3 mL SOC + 2 mL transformed Endura from two separate transformations). Pro-Tip Do not pipette...assumes familiarity with standard bacterial transformation and basic knowledge of the library being amplified...amplified. Please see our protocol on Bacterial Transformation . The generalized pooled library amplification...using a new vented falcon tube for every two transformations. Each of the four 14 mL Vented Falcon Tubes...repeatedly or mix when removing SOC containing transformed DNA-Endura from cuvette. Shake four 14 mL Vented...plates at 30 ℃ overnight. Plate 2.5 mL of the transformed cells on each of the eight bioassay plates (two...

  20. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...self-ligating recipient plasmid backbone. Transformation Transform your ligation reaction into your bacterial...from your transformation will give you the first indication as to whether your transformation worked. our...Digest of Plasmid DNA DNA Ligation Bacterial Transformation Summary The following technique can be used...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 should also verify that you are...
Showing: 401 - 420 of 457 results