The Ion AmpliSeq™ Direct FFPE DNA Kit enables the preparation of DNA from formalin-fixed, paraffin-embedded (FFPE) tissues for library construction using the Ion AmpliSeq protocol, without the need for deparaffinization or DNA purification. This kit is suitable for FFPE tissue sections up to 10 µm thick.
Features of the Ion AmpliSeq Direct FFPE DNA Kit include:
• Simple one-tube 2-step protocol that minimizes sample loss
• Efficient 30-minute workflow with only 10 minutes of hands-on time
• Ideal for FFPE DNA samples with limited DNA amount
• No deparaffinization of FFPE sections required
• No column or bead-based DNA purification required
• Ion AmpliSeq library preparation with as little as 1 ng input DNA
• Uracil-D-glycosylase treatment to remove deaminated cytosines is an optional step
Extraction of DNA from samples with limited DNA amount
Archived tissue samples can provide clinically relevant information, but it can be difficult to isolate the small amount of nucleic acid they contain. Deparaffinization of FFPE tissue sections and purification of the DNA in these samples can be a complicated, multi-step procedure requiring considerable hands-on time, and the associated sample-loss makes it an unsuitable approach for processing FFPE DNA samples with limited DNA amounts.
A simpler and faster method
The Ion AmpliSeq Direct FFPE DNA Kit provides a user-friendly protocol that eliminates the need for deparaffinization of FFPE tissue sections and purification of the DNA. From an FFPE section mounted on a glass slide, the area of interest is removed using Transfer Solution and pipetted into a PCR tube or 96-well plate. After the addition of Direct Reagent, the sample is incubated at 65°C for 15 minutes. The FFPE DNA is now ready for the Ion AmpliSeq™ Library Preparation protocol. The prepared FFPE DNA can be quantified using the Qubit™ HS DNA Quantitation Kit, if necessary. The simplicity of this protocol eliminates the need for complicated deparaffinization and DNA extraction procedures, and libraries can be prepared either manually or with the automated Ion Chef™ System.