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Genomic DNA Purification from Sample Applied to FTA Cards

Materials

FTA Card (Micro, Mini, or Classic Card); Indicating card for clear colorless samples

Harris Micro Punch or disposable Uni-Core Punch (1.2, 2.0, or 3.0 mm coring device, depending on sample and downstream application)

FTA Purification Reagent

TE-1 buffer (10 mM Tris-HCl, 0.1 mM EDTA, pH 8.0)*

* Note that this buffer contains 0.1 mM EDTA, not 1 mM EDTA.

Advance preparation

None

Protocol

1. Collect sample and dry

Apply liquid sample (125 µL per 25 mM circle), crushed tissue, or plant leaf to the FTA card and air dry for 1 h. Cells are lysed, proteins are denatured, pathogens are inactivated, and DNA is released to entangle in the fibers of the matrix.

Card must be completely dry before proceeding to punching and purification.

2. Punch

Cut a disc using a Harris Micro Punch or Uni-Core device and place in a PCR tube.*

3. Rinse the punch with FTA Purification Reagent

Wash with 200 µL of FTA Purification Reagent for 5 min at room temperature; perform three washes. Cell debris and PCR inhibitors are washed away.

4. Rinse the punch with TE-1 buffer

Wash with 200 µL of TE-1 buffer for 5 min at room temperature; perform two washes. FTA Purification reagent, which itself is a potent PCR inhibitor, is washed away.

TE with reduced EDTA content is used because EDTA may interfere with PCR.

5. Dry

Air dry the punch for 1 h at room temperature or for 15 min at 50 °C.

6. Perform PCR analysis

Add PCR Master Mix and perform PCR according to the protocol chosen.

* Guidelines: 1.2 mM punch for blood samples and samples with high DNA content; 2.0 mM for buccal cells, plant cells, and bacteria containing plasmid DNA; 3.0 mM to prepare DNA from FTA Cards using illustra tissue & cells genomicPrep Mini Spin Kit.

Materials
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