... 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...

...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...

... 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...

...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...

...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...

...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...

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

.... 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...

...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...

...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...

...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...

...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 ...

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

...mini-prep you will usually want to start 2 mL in a falcon tube, but for larger preps you might want to use...

...reaction in 1 hour. Using this ratio, you can calculate the minimal amount of enzyme for your reaction...

...50 μL of competent cells in a microcentrifuge or falcon tube. GENTLY mix by flicking the bottom of the ...

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

...base pairs for each varies, it is difficult to calculate this based on DNA concentration alone. One method...

...mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...

... 37 °C . Determine the absorbance at 590 nm . Calculate the average absorbance of the duplicate samples...