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Addgene

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

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We narrowed to 23 results for: ALC

Showing: 1 - 20 of 23 results

  1. Lentivirus ddPCR Titration

    Type
    Protocol
    ... forget to factor this into your calculations later (see calculation example below). Seed 300,000 cells... a CSV file which will be used to calculate the titer. Calculations In the experimental setup above, the...20 200 40 400 80 800 160 1600 To calculate the titers, first calculate the number of viruses per cellular...glutaGRO, Corning 25-015-CI) 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...over copies\ RPP30}$$ Use the viruses/genome to calculate the infectious titer: $$T = {V*C*D\over v}$$ Where...titer obtained from the appropriate dilutions to calculate the final infectious titer. Tips and Troubleshooting...green). To increase the accuracy of the titer, calculate an average of several dilutions. Figure 1: ddPCR...

  2. CRISPR Library Amplification

    Type
    Protocol
    ...new vented falcon tube for every two transformations. Each of the four 14 mL Vented Falcon Tubes should... mL of LB Agar per bioassay plate. 14 mL Vented Falcon Tubes (BD Biosciences, 352059) 3X LB Agar + Antibiotic...supports normal growth is expected to work. 50 mL Falcon Conical tubes (Fisher, 14-432-22) Reagent Preparation...Aliquot 3 mL SOC into each of four 14 mL Vented Falcon Tubes and have 1 mL SOC per electroporation readily...all liquid from cuvette and add to 14 mL vented Falcon Tube containing 3 mL SOC. Repeat for each of the...DNA-Endura from cuvette. Shake four 14 mL Vented Falcon Tubes at 30-37 ℃, 225 rpm for 1 hour. After the...

  3. Fluorescence Titering Assay

    Type
    Protocol
    ... of PBS. Calculate the fraction of fluorescent-positive cells in each well. When calculating titer, only...mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...remaining well, a cell count is required for calculating the titer. Incubate for 48–72 h. Gently aspirate...leading to underestimation of the true titer. Calculate the transduction units per mL (TU/mL) using either...

  4. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...CI) Heat-inactivated FBS 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...will be used to seed the 96-well plate. Sample Calculation First, use a hemocytometer to quantitate the ...well plate, make 10 mL of a 5 cell/mL solution. Calculate the total cells needed: Total cells needed: 10...

  5. Kit Free RNA Extraction

    Type
    Protocol
    ...Phenol 2 M Sodium Acetate pH 4 Chloroform/Isoamyl alcohol (49:1) 75% Ethanol RNase-free water or TE solution...by inversion. Add 0.2 mL of Chloroform/Isoamyl alcohol (49:1) and then shake vigorously by hand for 10...homogenized sample(s). Add 0.2 mL of Chloroform/Isoamyl alcohol (49:1) per 1 mL of TRIzol® used. Shake vigorously...

  6. AAV Production in HEK293 Cells

    Type
    Protocol
    ...Thermo Fisher, 12605010) 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...incubator for 24 h–36 h. Proceed with transfection: Calculate the amount of each plasmid needed to have a 1:... Therefore, for each plasmid we need: Sample Calculations RepCap: 0.08 μg/bp × 7,265 bp = 582.1 μg Volume...

  7. Centrifugation

    Type
    Protocol
    ... the two, use a conversion calculator or chart like the G Force calculator (Link opens in a new window...

  8. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    .... Use a (Link opens in a new window) ligation calculator to easily quantify how much vector and insert...vector end. (Link opens in a new window) Ligation calculators are easily found on the web. Just enter the concentration...

  9. Colony Formation Titering Assay

    Type
    Protocol
    ...mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...integration events decreases as the dilution increases. Calculate the transduction units per mL (TU/mL) as follows...

  10. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...Addgene. The values highlighted below in red were calculated using this plasmid, but will change if you use...use a different plasmid. Sample Calculation Size in bp of Addgene plasmid #59462 : 6208 bp Concentration...

  11. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3’) into this calculator we get a final Reverse Primer sequence of 5’-...base pairs for each varies, it is difficult to calculate this based on DNA concentration alone. One method...

  12. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...trendline for the standards. Google Sheets can calculate this for you as follows: In the Chart Editor ,...based on your sample’s HC + LC area. Example calculation: If the sample HC + LC Area is 18179.442 and ...

  13. DNA Quantification

    Type
    Protocol
    ...Calculate your DNA sample concentration and purity using Addgene’s DNA Quantification Protocol....

  17. Lentivirus Production

    Type
    Protocol
    ... Da Heat-inactivated FBS 1x PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...

  20. Western Blot

    Type
    Protocol
    ... 37 °C . Determine the absorbance at 590 nm . Calculate the average absorbance of the duplicate samples...
Showing: 1 - 20 of 23 results