Volume : 96 preps per kit.
The kit can be integrated with an automatic nucleic acid extractor, such as KingFisher, for high-thro µghput extraction experiments, or manually operated with a magnetic separation rack.
The purification system uses superparamagnetic nano-magnetic particles as the matrix, which can adsorb nucleic acids specifically thro µgh hydrogen bonds and static electricity under the condition of high concentration of leachate, while proteins and other non-specific impurities are removed by washing. Finally the nucleic acid are eluted with low salt buffer or RNase-free ddH2O. The purified nucleic acid can be used for various routine operations, including RT-PCR, RT-qPCR, fluorescence quantitative PCR (qPCR) and other downstream experiments.
Main components
The kit consists of the following components :
Component
Volume
Plate-A (Lysis Buffer/Mag Beads)
622µl/well
Plate-B (Wash Buffer)
600µl/well
Plate-C (Wash Buffer)
600µl/well
Plate-D (Elution Buffer)
50µl/well
Proteinase K
1mL × 2
Magnetic Head
1 pc
Prepare 1× PBS solution, pH 7.4, in case of need.
Storage conditions
The kit can be stored for 12 months at room temperature (15-25°C) under dry conditions.
Notes
1. Avoid repeated freezing-thawing of samples, otherwise the extracted viral RNA will be degraded and the extracted amount will decrease. The samples can be extracted immediately or stored at 4°C for testing for no more than 24 hours. Long-term storage can be placed at -20°C or -80°C.
2. All operating procedures, if not specified, are carried out at room temperature (15-25°C).
3. When using this kit, please wear lab coat, disposable latex gloves, disposable masks and use RNase-free consumables to avoid RNase pollution to the greatest extent. Prepare your own RNase-free pipette tips, etc.
4. Please read the instructions caref µLly before use and follow the instructions strictly. Clinical samples sho µLd be carried out in the µLtra clean table or biosafety cabinet.
5. There may be residual magnetic beads during elution, magnetic beads sho µLd be avoided as far as possible when pipetting samples.
6. The virus has a strong ability to infect, a variety of defense measures must be done before the operation. Proper disposal of samples and reagent materials, thoro µgh cleaning and disinfection of the operating table.
Sample preparation
A. Throat swab (with preservation solution), saliva : vortex vigorously for 30 sec, take 200 µL for experiment.
B. Plasma, serum and viral stock solution : prepare 10-200µl of plasma, serum or viral stock solution, if the initial amount is less than 200 µl, use PBS solution to make up to 200 µl.
C. Virus-infected tissue : prepare 10mg of virus-infected tissues to be ground with liquid nitrogen, and add 200µl of PBS solution to the ground tissues.
Protocol : Automatic Operation Process of 96 Deep-Well Plate
1. Take out the pre-packaged 96 deep-well plates.
2. Oscillate four angles of the Plate-A to make the Mag Beads suspended.
3. Open the sealing membranes of the 96 deep-well plates.
4. Add 20 µl Proteinase K and 200µl sample to each well of the Plate-A.
5. Place the Magnetic Head into the Plate-C.
6. Follow the instructions to put the 4 pre-packaged plates into the correct position of the machine.
7. Run the viral nucleic acid extraction program of magnetic bead method “gdsbio_normal_96.bdz”, or refer to the “Sheet of Program Design” below.
8. At the end of the automation process, in the Plate-D is the DNA/RNA solution, sealed with a sealing membrane and stored at -20°C or -80°C.
Sheet of Program Design
Plate
Operation Process
A
Mixing at 20~55°C for 10 min by vibrating, and the digested sample release DNA/RNA to the Mag Beads. Incubate for 2 min.Tansfer the beads to Plate-B.
B
Wash the beads by vibrating for 2 min, then tansfer the beads to Plate-C.
C
Wash the beads by vibrating for 2 min, dry the beads for 2 min, then tansfer the beads to Plate-D.
D
Elute the beads by vibrating for 2 min, then tansfer the beads back to Plate-C.
Suitable Instruments
Thermo KingFisher Flex96,