Buccal Swab （FTA Card）DNA Isolation Kit

Description

Recovering concentrated genomic DNA from a single buccal swab or a limited FTA blood spot card sample can be challenging when only a small amount of starting material is available. Conventional purification methods may also require organic extraction or time-consuming alcohol precipitation, adding handling steps before downstream research.

The Buccal Swab & FTA Card DNA Isolation Kit provides a spin-column genomic DNA purification workflow for one buccal swab or 1–3 FTA blood spot card punches approximately 3 mm in diameter. The procedure combines Foregene Protease digestion with a DNA-only silica membrane spin column.

According to the instruction manual, genomic DNA can be extracted in approximately 80 minutes. The 15–100 μL elution range allows the Buffer EB volume to be selected according to the needs of the downstream experiment. Buffer EB should not be used below 15 μL because this may affect DNA yield.

The buccal swab and FTA card DNA extraction procedure does not require phenol or chloroform extraction and does not require isopropanol or ethanol precipitation. Absolute ethanol is still required for Buffer WB preparation and during sample processing.

The kit can process one sample or multiple samples, using a separate DNA-Only Column for each purification.

For research use only.

Specifications

Kit Components

Safety note: Buffer ST2 and Buffer PW contain irritating desalted salt. Wear gloves and take appropriate protective measures during handling.

Required Materials and Equipment

The operating procedure requires the following materials or equipment in addition to the components listed in the kit:

• Absolute ethanol

• Clean 2 mL centrifuge tubes

• Clean 1.5 mL centrifuge tubes

• Microcentrifuge capable of 12,000 rpm, approximately 13,400 ×g

• Vortex mixer

• Metal bath or water bath capable of maintaining 65°C

• 37°C water bath when buffer precipitate must be dissolved

• Pipettes and compatible pipette tips for the specified liquid volumes

Buffer preparation: Add 60 mL of absolute ethanol to Buffer WB supplied with the DE-05811 kit before use. The protocol also requires 200 μL of absolute ethanol during each sample purification.

Applications

Buccal Swab Genomic DNA Extraction

The kit is intended for genomic DNA purification from one buccal swab per purification. The cotton portion of the sample swab is placed into a clean 2 mL centrifuge tube and subjected to Foregene Protease digestion before spin-column purification.

FTA Blood Spot Card DNA Extraction

For FTA card DNA extraction, the instruction manual recommends using 1–3 blood spot card punches approximately 3 mm in diameter for each purification. If the blood card was not stored under suitable conditions, the manual states that the amount of sample material may be increased.

Low-Volume Genomic DNA Elution

The 15–100 μL Buffer EB range supports genomic DNA elution at different volumes. Buffer EB should not be used below 15 μL because this may affect DNA yield. Increasing the elution volume within the recommended range may improve DNA recovery.

Downstream Research Preparation

The purified genomic DNA is intended for downstream detection or research experiments. Researchers should determine compatibility with the requirements of their specific downstream workflow.

Workflow Overview

1. Prepare the sample. Place the cotton portion from one buccal swab or 1–3 FTA blood spot card punches approximately 3 mm in diameter into a clean 2 mL centrifuge tube.

2. Add Buffer ST1. Add 300 μL Buffer ST1 and mix by vortexing.

3. Add Foregene Protease. Add 20 μL Foregene Protease, vortex for 5 seconds and briefly centrifuge.

4. Perform the first incubation. Incubate at 65°C for 1 hour and vortex for 5 seconds at 15-minute intervals.

5. Add Buffer ST2 and Linear Acrylamide. Add 310 μL Buffer ST2 followed by 4 μL Linear Acrylamide. Vortex for 5 seconds and briefly centrifuge.

6. Perform the second incubation. Incubate at 65°C for 10 minutes, mixing once every 3 minutes.

7. Prepare the lysate for binding. Remove the buccal swab when applicable, add 200 μL absolute ethanol and vortex for 5 seconds. Flocculent precipitate may appear.

8. Bind the DNA. Transfer the solution and any flocculent precipitate to the DNA-Only Column. Centrifuge at 12,000 rpm, approximately 13,400 ×g, for 1 minute.

9. Wash with Buffer PW. Add 500 μL Buffer PW and centrifuge at 12,000 rpm, approximately 13,400 ×g, for 1 minute.

10. Wash twice with Buffer WB. Add 700 μL Buffer WB and centrifuge for 1 minute. Repeat the Buffer WB wash once.

11. Dry the membrane. Centrifuge the empty spin column at 12,000 rpm, approximately 13,400 ×g, for 1 minute.

12. Elute the genomic DNA. Apply 15–100 μL Buffer EB preheated to 65°C to the center of the membrane. Incubate at room temperature for 5 minutes and centrifuge for 1 minute.

Refer to the complete instruction manual before beginning the procedure. This overview does not replace the complete operating instructions.

Important Operating Notes

• Use one buccal swab for each purification.

• Use a separate DNA-Only Column when processing multiple buccal swabs.

• Use 1–3 FTA blood spot card punches approximately 3 mm in diameter for one purification.

• Keep the sample immersed in the lysis buffer during sample lysis.

• If the sample adheres to the tube cap or inner wall, collect it by brief centrifugation.

• Inspect Buffer ST1, Buffer ST2 and Buffer PW for precipitation before use.

• If precipitation is present, warm the affected buffer at 37°C and mix thoroughly until dissolved.

• Confirm that absolute ethanol has been added to Buffer WB before use.

• Do not add RNase to any buffer.

• Do not use less than 15 μL Buffer EB for elution.

• Apply Buffer EB to the center of the membrane rather than the press ring.

• During the binding step, transfer both the solution and any flocculent precipitate to the DNA-Only Column.

Elution Guidance

Use 15–100 μL Buffer EB preheated to 65°C. Apply the buffer to the center of the spin-column membrane, leave the column at room temperature for 5 minutes and centrifuge at 12,000 rpm, approximately 13,400 ×g, for 1 minute.

Within the recommended elution range, increasing the Buffer EB volume may improve DNA yield. When increased DNA concentration is required, the first eluate may be reapplied to the spin column and centrifuged again for 1 minute.

Storage and Stability

• Store the complete kit under dry conditions at 15–25°C for up to 12 months.

• For longer storage, the kit may be stored at 2–8°C.

• The instruction manual states that Foregene Protease remains active at room temperature for 3 months.

• Storage of Foregene Protease at 4°C is recommended for improved activity and stability.

• Do not store Foregene Protease at −20°C.

Low-temperature storage may cause solution precipitation. Before use, allow the kit solutions to return to room temperature. If necessary, warm the affected solution in a 37°C water bath for 10 minutes, mix thoroughly and confirm that the precipitate has dissolved.

Documents

Buccal Swab & FTA Card DNA Isolation Kit Instruction Manual, Version 1.0

Frequently Asked Questions