PCR Metabarcoding Simulator

Assay Setup

Template A — N0 (molecules)

Template B — N0 (molecules)

Template A — Amplification Efficiency

E_A0.90

Template B — Amplification Efficiency

E_B0.60

Total Primer Mix [nM]

Volume [µL]

Cycles

Forward Primer & Variants

Primer Sequence (IUPAC: R=AG, Y=CT, W=AT, S=CG, K=GT, M=AC, B=CGT, D=AGT, H=ACT, V=ACG, N=ACGT)

-

Variants A (all that bind A)

∩

Overlap A∩B

∩

Variants B (all that bind B)

-

Each copy binds one of its variants at random. A splits its demand across exclusive + overlap variants. Within the overlap, A and B compete in proportion to copy number. Once the overlap is exhausted, B stops, while A continues using its exclusive variants.

Model

Calculation Used

-

Amplification Stop (Primer Exhausted)

Template A

-

Template B

-

Absolute Copies vs. Cycles

Relative Reads vs. Cycles

Results Table

Cycle	Copies A	Gain A	Copies B	Gain B	Proportion A	Proportion B