HiPure FFPE DNA Kit

How to read this note

1. Workflow structure
This workflow separates sample pretreatment from the shared downstream purification steps. It is intended as a practical companion to the product manual rather than a replacement for the official protocol. The downstream steps follow a silica membrane-based column purification workflow.

2. Time interpretation
Protocol times stated in the product manual are retained where applicable. Steps without explicit timing are estimated for an experienced operator. This workflow separates active handling time from prolonged digestion time. For short protocol ranges, the timeline uses the midpoint. For long sample-dependent ranges, the timeline shows the shortest reasonable workflow path, while the broader range is explained in the note.

3. Workflow characteristics
This workflow is designed for FFPE DNA recovery after paraffin removal, protease-assisted lysis and thermal reversal of formaldehyde-induced crosslinking. Compared with routine tissue DNA extraction, the defining upstream feature is the 90°C reversal step, which improves DNA accessibility for silica binding but can also increase fragmentation if prolonged excessively.

4. Practical considerations
In practical use, complete digestion is usually more important than shortening the pretreatment stage. Dense, heavily fixed or older FFPE material may require extended 56°C incubation beyond the 1 h minimum shown in the timeline. If RNA-free DNA is required, RNase A treatment can be added after cooling from the 90°C step. Avoid overloading the column with incompletely lysed material, as this may reduce recovery or contribute to clogging.