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Restriction Digest Calculator

Single and double digests: µL of DNA, enzyme, buffer and water for the amount of DNA you want to cut. The same math as the Excel workbook, live in your browser.

What this is for: The µL of enzyme, buffer, DNA and water for a single-enzyme digest at a chosen units-per-µg rate.

The formula: enzyme (µL) = DNA (µg) × units per µg ÷ stock concentration (U/µL)

How to use:

  1. For a single digest, use section A: enter the DNA concentration (ng/µL), DNA amount wanted (µg), final reaction volume (µL) and enzyme volume (µL).
  2. For a double digest, use section B: enter the same values plus the volume (µL) of each of the two enzymes.
  3. Read the DNA to pipette (µL), the automatic 10X buffer volume (µL) and the water to total (µL) for that section.
  4. Keep the total enzyme volume at or below 10% of the reaction (glycerol) and check buffer compatibility for double digests.

A. Single Digest

Buffer volume is automatic (total / 10 for a 10X buffer).

DNA to pipette (µL)
10X buffer (µL)
Water to total (µL)

B. Double Digest

Two enzymes, same total volume.

DNA to pipette (µL)
10X buffer (µL)
Water to total (µL)
Prefer to work offline or keep a copy? The download link is right below.
Download the Restriction Digest Calculator workbook (.xlsx)
Notes & limits (from the workbook): Buffer volume is automatic (total / 10 for a 10X buffer). Keep total enzyme volume ≤ 10% of the reaction (glycerol) and check buffer compatibility for double digests.