Figure 1.
Simulation diagrams of emulsion PCR amplification of a random DNA library.
Different compartment contained different templates marked with different colors, and some compartment are empty.
Figure 2.
Conventional PCR amplification of a DNA random library versus emulsion PCR.
PAGE electropherograms of PCR reaction mixtures for amplification of a random DNA library with conventional PCR (A) and emulsion PCR (C). Microfluidic chip electrophoresis electropherograms of PCR reaction mixtures for amplification of a random DNA library with conventional PCR (B) and emulsion PCR (D). Dynamics of product and by-product concentrations in conventional PCR and emusion PCR (E). Conventional PCR amplification was performed with cycles ranging from 10 to 32; the template concentration was 0.01 pmol/ml; the primer concentration was 0.4 µmol/L; and the concentration of Taq DNA polymerase was 0.05 U/µl. Emulsion PCR amplification was performed with cycles ranging from 10 to 40; the template concentration was 0.01 pmol/ml; the primer concentration was 0.4 µmol/L; and the concentration of Taq DNA polymerase was 0.125 U/µl.
Figure 3.
Influence of the number of template molecules.
Panel A indicates the PAGE electropherograms of PCR reaction mixtures for amplification of a random DNA library with different amounts of template molecules. Panel B indicates the concentration of dsDNA products and by-products obtained from microfluidic chip electrophoresis. Emulsion PCR amplification was carried out for 35 cycles; the annealing temperature was 65°C; the primer concentration was 0.4 µmol/L; and the concentration of Taq DNA polymerase was 0.125 U/µl.
Figure 4.
Influence of the annealing temperature.
The concentration of the dsDNA products and by-products was obtained from microfluidic chip electrophoresis. Emulsion PCR amplification was carried out for 35 cycles; the number of template was 0.01 pmol/ml; the primer concentration was 0.4 µmol/L; and the concentration of Taq DNA polymerase was 0.125 U/µl.
Figure 5.
Influence of DNA polymerase concentrations.
Emulsion PCR amplification was carried out for 35 cycles; the number of templates was 0.01 pmol/ml; the primer concentration was 0.4 µmol/L; and the annealing temperature was 65°C.
Figure 6.
Influence of primer concentrations.
Emulsion PCR amplification was carried out for 35 cycles; the number of templates was 0.01 pmol/ml; the concentration of Taq DNA polymerase was 0.125 U/µl; and the annealing temperature was 65°C.
Figure 7.
Conventional PCR was carried out for 1 and 30 cycles with 5 µg/ml template. 1 and 6: PCR mixture only include template. 2 and 7: PCR mixture include all except primer. 3 and 8: PCR mixture include all except dNTP. 4 and 9: PCR mixture include all except DNA polymerase. 5 and 10: PCR mixture include all the masteries.