Quality Molecular Bio Reagent Cell Free cf Dna Extraction Kit
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- Supplier: Foregene Co. Ltd.
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Product Overview
Description
Quality Molecular Bio Reagent Cell Free cf Dna Extraction Kit
Description:
The FOREGENE Cell-free DNA Isolation Kit is designed for isolating circulating cell-free DNA (cfDNA) from human plasma, serum, or urine samples. The kit can quickly and conveniently isolate cfDNA in three steps: lysis/binding, washing and elution. It uses specially designed silica-coated superparamagnetic beads with a unique lysis/binding system to recover circulating nucleic acids by hydrogen bonding and electrostatic forces, without adsorbing proteins and other impurities. It is suitable for high-throughput automated extraction work- stations in preparation for any downstream analysis using PCR or NGS.
Kit content and storage:
Table 1 FOREGENE Cell-free DNA Isolation Kit
Contents | Reagent | Amount(25 T) | Amount(50 T) | Amount(100 T) | Storage |
Box1 |
Buffer AL |
42 mLx1 bottle |
89 mLx1 bottle |
189 mLx1 bottle | 10-30°C |
Buffer AW1 | 38 mLx1 bottle | 83 mLx1 bottle | 165 mLx1 bottle | 10-30°C | |
Buffer AW2 | 15 mLx1 bottle | 33 mLx1 bottle | 66 mLx1 bottle | 10-30°C | |
Elution | 3 mLx1 bottle | 6 mLx1 bottle | 12 mLx1 bottle | 10-30°C | |
Box2 |
Magnetic Beads A |
3.2 mLx1 bottle |
6.5 mLx1 bottle |
13 mLx1 bottle |
2-8°C |
Proteinase K |
5 mLx1 bottle |
10.5 mLx1 bottle |
21 mLx1 bottle |
2-8°C |
Notes:
[1] Do not mix components from different batches of kits.
[2] Buffer AL may form precipitates, which does not affect its function. If precipitation is visible, please place the reagent bottle in a 56°C water bath for 10-20 min until the precipitate dissolves. Then mix thoroughly before use.
[3] Do not freeze the Magnetic Beads A.
Required equipment and materials not supplied:
Table 2 Materials required for cfDNA isolation
Item | Note |
Equipment | |
Bench-top centrifuge | Rotation speed not lower than 10,000 rpm/min |
Handheld Mini Centrifuge | / |
Vortex | / |
Thermostatic metal bath | / |
Magnetic Separation Device | 1.5 mL, 5mL |
Micro-Pipettes | 1 mL, 200 μL, 20 μL, 10 μL |
Reagents | |
Absolute Ethanol (>80%) | Analytical Grade |
Isopropanol | Analytical Grade |
Consumables | |
Centrifuge tubes | 5 mL, 1.5 mL |
Aerosol-resistant pipette tips | 1 mL, 200 μL, 20 μL, 10 μL |
Important points before starting
Avoid repeatedly freezing and thawing samples, which may result in low DNA quality and yield.
All reagents and samples need to equilibrate to room temperature (10°C ~30°C) before use.
Please use the recommended consumables for automated or manual operations.
Please read the manual carefully before the experiment.
Things to do before starting
1)Prepare the buffers according to Table 3. Add isopropanol to Buffer AL and add absolute ethanol to Buffer AW1 and Buffer AW2. Mix well, label and store at room temperature.
2)If precipitates are visible in Buffer AL or Buffer AW1, dissolve by placing bottle in a 56 °C water bath. Shake well to mix before use.
3)Centrifuge pre-cooling to 4°C.
4)Remove the magnetic bead from 2-8℃ and re-warm it at room temperature for 30 min, vortex it well before use.
5)Sample pretreatment: Plasma samples were centrifuged at 4°C for 10 min at 16000 g, and the supernatant was taken from 0.5-4 mL.
6)Prepare the necessary reagents and consumables according to Table 4. The materials needed for a 0.5- 4mL sample is shown. Volumes can be adjusted accordingly.
Table 3 Buffer preparation: addition of isopropanol / ethanol
Reagent | 2 mL×25 T | 2 mL×50 T |
2 mL×100 T |
Add isopropanol to Buffer AL | 8 mL | 17 mL | 36 mL |
Add ethanol to Buffer AW1 | 38 mL | 83 mL | 165 mL |
Add ethanol to Buffer AW2 | 60 mL | 132 mL | 264 mL |
Table 4 Consumables and Reagents for 0.5-4 mL Sample
Sample Volume | 0.5 mL | 1 mL | 2 mL | 4 mL |
Consumables | Volume | Volume | Volume | Volume |
Centrifuge Tube | 1.5 mL | 5 mL | 5 mL | 10 mL |
Reagents | Volume | Volume | Volume | Volume |
Proteinase K | 50 μL | 100 μL | 200 μL | 400 μL |
Buffer AL | 0.5 mL | 1 mL | 2 mL | 4 mL |
Magnetic Beads A | 32 μL | 62.5 μL | 125 μL | 250 μL |
Buffer AW2 | 1.5 mL | 1.5 – 3 mL | 3 mL | 3 mL |
Buffer AW2 | 1.5 mL | 1.5 – 3 mL | 3 mL | 3 mL |
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