PCR Master Mix

Build 2x master mix setup tables with overage, copy-friendly TSV output, and CSV export.

For a 2x master mix: half the final reaction volume is premix. Include controls in the sample count and verify the reagent protocol.

Inputs

sample Countextra Percentagereaction Volumetemplate Volumeforward Primer Volumereverse Primer Volume

Output

Total reactions with overage: 13.2

Component	Per reaction (uL)	Total (uL)
Nuclease-free water	8.5	112.2
2x master mix	12.5	165
Forward primer	1	13.2
Reverse primer	1	13.2
Template DNA	2	26.4
Total	25	330

Method and worked example

Equivalent reactions = samples × (1 + overage / 100). A 2x premix takes half of the final reaction volume. Water per reaction is the final volume minus premix, primers, and template. Multiply each per-reaction volume by equivalent reactions to obtain the table totals; fractional equivalent reactions are retained.

For 12 samples with 10% overage, prepare volumes for 13.2 equivalent reactions. At 25 µL per reaction, 2x premix is 12.5 µL per reaction, or 165 µL total. With 1 µL of each primer and 2 µL template, water is 8.5 µL per reaction, or 112.2 µL total.

Assumptions and limits

This worksheet assumes a 2x premix. Follow the reagent manufacturer's protocol for buffer, enzyme, primer, and template requirements. Template DNA is included in volume accounting, but different sample templates should generally be added separately; the total row is not an instruction to pool templates. Include controls in the sample count and choose enough overage for your pipetting workflow.