FireGene Plasma Cell-Free DNA Extraction Kit (Magnetic Bead Method)

Intended Use

Used for nucleic acid extraction, enrichment, and purification. The processed product is intended for research use. Suitable for extracting high-quality cell-free DNA from the plasma of humans and other mammals.

Test Principle

This kit uses silica-coated, superparamagnetic nanobead purification technology suitable for purifying and recovering cell-free DNA from plasma. The kit can be used for both manual and automated extraction. Under high-salt conditions, cell-free DNA binds to the surface of the silica-coated magnetic beads. After washing, DNA is eluted from the beads using the washing liquid. The purified cell-free DNA is of stable, reliable quality and can be used in downstream applications such as NGS, genotyping, and qPCR.

Specifications

Product Name	FireGene Plasma Cell-Free DNA Extraction Kit (Magnetic Bead Method)
Catalog No.	FG-FG1102D
Package Size	96 reactions/kit
Purification Technology	Silica-coated superparamagnetic nanobeads
Extraction Format	Compatible with both manual and automated extraction
Sample Type	Plasma (human and other mammals)
Downstream Applications	NGS, genotyping, qPCR
Intended Use	Research Use Only (RUO)
Transport Conditions	Ambient temperature, no more than 10 days
Storage Conditions	Store Proteinase K and Magnetic Bead Suspension C at 2–8°C; store other components at ambient temperature, protected from light
Shelf Life	12 months

Main Components (96 reactions)

Component	96 rxns
Lysis Buffer 1	50 mL
Lysis Buffer 2	340 mL
Proteinase K	17 mL
Magnetic Bead Suspension C	9 mL
Washing Solution C1*	110 mL
Washing Solution C2*	45 mL
Washing Liquid C	65 mL

*Before first use, add the amount of isopropanol indicated on the label to Washing Solution C1, and the amount of absolute ethanol indicated on the label to Washing Solution C2; mix well by shaking before use. Since ethanol and isopropanol readily evaporate onto the bottle walls, shake Washing Solutions C1 and C2 well before each subsequent use.

Compatible Instruments

Validated nucleic acid extraction instruments for plasma extraction. 

Sample Requirements

Plasma.

Required but Not Provided: Reagents, Equipment, and Consumables

Reagents to be supplied by the user:

Reagent Name	Concentration
Absolute Ethanol	≥ 99.7%
Isopropanol	≥ 99.7%

Equipment and consumables to be supplied by the user: Thermostatic shaker/mixer, vortex mixer, mini centrifuge, magnetic separation rack for microtubes or a nucleic acid extraction instrument (with its compatible consumables), 1.5 mL nuclease-free centrifuge tubes, 15 mL nuclease-free centrifuge tubes, high-speed centrifuge, etc.

Note: To ensure optimal reagent performance, mixing reagents from different lots is not recommended.

Protocol Overview — Manual Extraction (4 mL plasma example)

1. Lysis: In a 15 mL centrifuge tube, add 160 µL Proteinase K, 4 mL sample, and 400 µL Lysis Buffer 1, in that order, and mix by vortexing. Incubate at 60°C in a water bath for 20 minutes. To avoid inactivating Proteinase K, add components in the order Proteinase K → sample → Lysis Buffer 1.
2. Binding: Add 3.2 mL Lysis Buffer 2, 1.8 mL isopropanol, and 80 µL Magnetic Bead C, and mix by vortexing. Bind at room temperature for 10 minutes, then place on the magnetic rack and discard the supernatant once clear.
3. Wash 1 & 2: Add 1 mL Washing Solution C1, vortex 20 s, place on magnetic rack, discard supernatant.
4. Wash 3 & 4: Add 1 mL Washing Solution C2, vortex 20 s, place on magnetic rack, discard supernatant.
5. Removing residual liquid: Briefly centrifuge, place on magnetic rack, aspirate residual liquid, and air-dry at room temperature for 5 minutes.
6. Elution: Add 40 µL Washing Liquid C, mix by pipetting, let stand at room temperature for 10 minutes (or in a metal bath at 25–30°C, 1,600 rpm, for 10 minutes).
7. Transferring cfDNA: Briefly centrifuge, place on the magnetic rack, and transfer the supernatant to a new nuclease-free 1.5 mL tube. Store at -20°C/-80°C.

Binding system for different plasma volumes:

Plasma Volume	1 mL	2 mL	4 mL	10 mL
Proteinase K	40 µL	80 µL	160 µL	400 µL
Sample Volume	1 mL	2 mL	4 mL	10 mL
Lysis Buffer 1	100 µL	200 µL	400 µL	1 mL
Total Volume	1.14 mL	2.28 mL	4.56 mL	11.4 mL

Protocol Overview — Automated Extraction (4 mL plasma example)

Lysis step is identical to the manual procedure. Separation and purification are then carried out across wells as follows:

Well No.	Function	Reagent / Sample
Well 1	Binding	3.2 mL Lysis Buffer 2 + 1.8 mL isopropanol + 4.56 mL lysed sample + 80 µL magnetic beads
Well 2	Washing solution C1	1 mL Washing solution C1
Well 3	Washing solution C2	1 mL Washing solution C2
Well 4	Washing solution C2	1 mL Washing solution C2
Well 5	Bead waste removal	500 µL Washing liquid C (used to discard waste beads)
Well 6	Elution	Add 50 µL Washing liquid C

Precautions

• Before use, add the amount of isopropanol indicated on the label to Washing Solution C1, and the amount of absolute ethanol indicated on the label to Washing Solution C2.
• Check Lysis Buffer 1 and Lysis Buffer 2 for precipitate or crystals before use. If present, dissolve in a 56°C water bath, then mix by shaking or inverting the bottle — using the buffer after the precipitate has dissolved does not affect performance.
• Before use, remove Magnetic Bead Suspension C from the 4°C refrigerator and let it equilibrate at room temperature for at least 10 minutes, then mix well by vortexing.
• For plasma separation from whole blood: centrifuge at 1,600 × g for 10 minutes and collect the supernatant, then centrifuge the supernatant at 16,000 × g for 10 minutes and use the resulting supernatant as the sample.