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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 29 results for: POR C

Showing: 21 - 29 of 29 results

  1. Gibson Assembly Protocol

    Type
    Protocol
    ... Cepko, 2020). Incubate the mix for 1 hour at 50 °C or follow manufacturer's instructions. You can purchase...Gibson DG, Smith HO, Hutchison CA, Venter JC, Merryman C. (2010). Chemical synthesis of the mouse mitochondrial....1515 (Link opens in a new window) Rabe BA, Cepko C. (2020). A Simple Enhancement for Gibson Isothermal...other. Phusion High-Fidelity DNA Polymerase - incorporates nucleotides to “fill in” the gaps in the annealed... product by restriction digest . Sequence the important regions of your final plasmid, particularly the...

  2. AAV ddPCR Titration

    Type
    Protocol
    ...parameters. Cycling Step Temperature (°C) Time (min) Ramp Rate (°C/sec) # Cycles Denaturation 95 10 2 1 ...sample: $$T = {R*C*D*1000\over V}$$ Where: T = Titer, GC/mL R = Reaction volume (20 µL) C = Copies/µL D ...Bio-Rad, 1863051 8-strip PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 48-well...place the PCR plate with the foil onto the metal support block. Place the block in the plate sealer and ... reading. When the droplet reading is complete, export the data from all wells as a CSV file and use the...

  3. Fluorescence 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 can promote...it can be inactivated in the lab by heating to 56 °C for 30 min. Considerations Before You Start The health...CI) Heat-inactivated FBS Polybrene (10mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium...

  4. CRISPR Library Amplification

    Type
    Protocol
    ...media at 37 °C (for at least 15 minutes). Prewarm 3X LB Agar + Antibiotic plates at 37 °C. Prewarm 8X ...Chill a box of 200 µL micropipette tips in a -20 °C freezer. Aliquot 3 mL SOC into each of four 14 mL ... mix. Electroporate cells (one at a time for a total of eight electroporations): Electroporator Conditions...and have 1 mL SOC per electroporation readily available for post-electroporation recovery of cells. Ensure...if the recombinant band makes up a significant proportion of the DNA pool. Last Update: August 17, 2023...limited. Equipment Table top centrifuge BioRad Electroporator (MicroPulser TM , Bio-Rad 1652100) Reagents.../or increasing the skewness of the pool. 8 electroporation cuvettes (BioRad, Micropulser, 0.1 cm ) 20 ...

  5. Colony Formation Titering Assay

    Type
    Protocol
    ...inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS can promote...it can be inactivated in the lab by heating to 56 °C for 30 min. Considerations Before You Start The health...CI) Heat-inactivated FBS Polybrene (10 mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium...

  6. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...standard Before Starting Warm the hot plate to 100 °C. Thaw IgG standard and prestained protein ladder on...microcentrifuge. Heat the samples for 10 min at 100 °C in a heat block. Spin the sample briefly in the microcentrifuge...IgG standards in Google Sheets. Open exported CSV file. Select import. Choose file. Format as needed. Select...that sharing the full details of our protocols supports reproducibility and accelerates science. Here,...information is solely for transparency intended to support reproducibility in science. Last Update: June 14...intensity of the protein bands to the overall lane. Import the gel image into ImageJ. Select File . Select... each peak by selecting the center of the peak. Export results as a csv file. In the results table, select...

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

    Type
    Protocol
    ...longer period of time; b) using a wider gel comb; or c) loading less DNA in the well. How do you get better...around the band as possible. To do so, it is often important to take the excised band, lay it down on the UV...sides with the razor blade. This is especially important during the DNA purification step, as many kits... fragment. The weight of the gel is directly proportional to its liquid volume and this is used to determine...manufacturer's instructions. Note: It is usually important to determine the concentration of the DNA that...

  8. Water Bath Protocol

    Type
    Protocol
    ... refrigerators to achieve a temperature between 4°C and room temperature. This general protocol suits ...on the surfaces of the water bath as the water evaporates from the tub. Disinfectant may be added to prevent...cover on the top of the water bath to prevent evaporation and maintain the desired temperature. This also...

  9. Ligation Independent Cloning

    Type
    Protocol
    ...software to ensure a melting temperature between 50-60°C for your PCR primers. Step 2: Linearize Vector In ...provided by your polymerase manufacturer. It is very important to remove all free nucleotides from your PCR product...
Showing: 21 - 29 of 29 results