Nucleic Acid Extraction Kit (Magnetic Bead Method)

Intended Use

This product is intended use for the extraction,enrichment and purification of nucleic acids. The extraction is intended use for clinical in vitro diagnosis.

Principle

High-quality nucleic acids can be separated and purified from complex samples by using a unique buffer system composed of lysis solution, washing solution A and washing solution B with separation function. The unique embeded magnetic beads have strong affinity with nucleic acids under certain conditions. The magnetic beads release the absorbed nucleic acids when the conditions change. This kit can extract nucleic acids with high purity and stable quality, which is suitable for the sensitive downstream applications, such as Real-time qPCR.

Storage And Stability

Store the kit at 15-30℃ and protected from light for 6 months.

Transporte at room temperature.

Production date and expiration date: See the label for details.

Applicable Devices

Manual operation: centrifuge, constant temperature mixer, 2mL magnetic rack (magnetic separator).

Specimen Requirements

1. Specimen Type

 saliva.

2. Specimen collection

Rinse mouth with clean water 30 min before collection (it is forbidden to collect immediately after rinsing), press the tip of your tongue against the root of the maxillary or mandibular teeth to enrich saliva, and gently spit saliva into the oval funnel until the liquid saliva (non-bubble) reaches the filling line height of 2 mL. Carefully unscrew the funnel from the collection tube (pay attention to make the collection tube stand upright during operation). Tighten the red cap and seal the collection tube. Check whether the sample collection tube is in good condition, and whether there is any sample overflow such as cracked tube and open cover. Discard the used funnel.

Test Procedure

1. Sample pretreatment

Centrifuge the saliva sample at 3000 rpm at 4℃ for 10 min, and collect the supernatant in a new centrifuge tube (be careful not to transfer the precipitate); The centrifuge tube was centrifuged at 13000 rpm at 4℃ for 2 min, and the collected supernatant could be extracted from the sample nucleic acid.

2. Manual extraction

2.1 sample cracking

Add 300 μL sample (the sample needs to be balanced to room temperature), 20 μL Proteinase K solution and 500 μL Lysis Buffer to 1.5 mL centrifuge tube, vortex for 10 seconds, and then place it on a constant temperature mixer at 80℃ and 1200 rpm for 4 min.

2.2 nucleic acid adsorption

Add 10 μL of Magnetic Beads Buffer to the centrifuge tube, vortex for 10 seconds, and then place it on a constant temperature mixer at room temperature and 1200rpm for 4 min.

   Place the centrifuge tube on the magnetic rack, and after the magnetic beads are completely absorbed, carefully absorb all the liquid.

2.3 Sample rinse 1

Add 500 μL of Washing Buffer A into the centrifuge tube, vortex for 10 seconds, and then place it on a constant temperature mixer at room temperature and 1200rpm for 2 min.

Place the centrifuge tube on the magnetic rack, and after the magnetic beads are completely absorbed, carefully absorb all the liquid.

2.4 Sample rinse 2

   Add 500 μL of Washing Buffer B into the centrifuge tube, vortex for 10 seconds, and then place it on a constant temperature mixer at room temperature and 1200rpm for 2 min.

Place the centrifuge tube on the magnetic rack, and after the magnetic beads are completely absorbed, carefully absorb all the liquid.

2.5 Nucleic acid elution

Dry the centrifuge tube for 2-5 min to ensure that there is no water residue.

Add 50 μL of Elution Buffer into the centrifuge tube, vortex for 10 seconds, and then place it on a constant temperature mixer at 56℃ and 1200rpm for 5 min.

Place the centrifuge tube on the magnetic rack, and after magnetic beads are adsorbed, collect the nucleic acids solution in a new centrifuge tube, and store at -20℃, or at -80℃ for long time.