Viral Swab/Saliva Nucleic Acid Extraction Kit (Magnetic Beads) 200 Preps

Genesis Swab/Saliva Viral DNA/RNA Extraction Kit (Magnetic Beads)

200 Preps

Description

Our Swab/Saliva Viral DNA/RNA Extraction Kit (Magnetic Beads) is a product specially designed for King Fisher and other nucleic acid extraction instruments. It is a small volume purification kit suitable for the extraction of various viral RNA/DNA from plasma, cell-free body fluids, viral stock solutions and infected tissues. The kit adopts the magnetic particle purification technology based on superparamagnetism, which can minimize the risk of cross-contamination and improve the sensitivity and accuracy of detection. The operation time of the instrument is only about 30 minutes. It can also be used for manual operation.

Main Components

Our kit consists of the following components:

Reagent	Amount	Component Description
Buffer MVL	65 ml × 2	Provides environment for lysing and binding to the magnetic beads
Buffer MW	20 ml × 3	Removes residual proteins and other impurities
Mag Beads	4.5 ml	Adsorbs viral nucleic acid
DEPC-Water	12 ml	DEPC - treated water, RNase - free
DS Carrier	450 μl	Captures trace nucleic acid
Proteinase K	1 ml × 4	Lyses proteins bound to nucleic acids

Storage Conditions

Store DS Carrier and Proteinase K at -20°C, Mag Beads and DEPC-Water at 2-8°C, others at room temperature (RT, 15-25°C), and transport at RT.

Notes

1. When using this kit, please wear a lab coat, disposable latex gloves, disposable masks to protect you from the reagents, and to protect the nucleic acid from nucleases that are present on the skin. The microcentrifuge tubes and pipette tips should be autoclaved and free of DNase and RNase.

2. Before using, vortex the Mag Beads well to ensure that the beads are fully resuspended. Mag Beads cannot be frozen.

3. For refractory samples, the lysis condition can be set to 55°C for 10 minutes.

4. Please check whether there is crystal precipitation in the Buffer MVL. If there is crystal precipitation, place it at room temperature or 37°C until the crystal is dissolved. Mix it before use.

5. In order to save time, Proteinase K, Mag Beads and DS Carrier can be pre-mixed, and the mixture can be placed at 2~8°C for 2 days. Mix 10 to 20 times to disperse the beads before using. Due to the

inhibition of Buffer MVL on Proteinase K, samples should be added as soon as possible after the addition of Buffer MVL.

6. This kit can be adapted to all the automatic instruments based on magnetic beads. If it is used for the first time, the operation procedure can be simulated through the empty plate, and then the samples can be added for extraction after it is accurate.

Before use

Add 80 ml of 100% ethanol to Buffer MW (20 ml), and store at RT.

Vortex the Mag Beads well to ensure that the beads are fully resuspended.

Prepare 1X PBS solution, pH 7.4 in case needed.

Sample Preparation

A. Throat swab (with preservation solution) or saliva: vortex vigorously for 30 sec and take 200 ul for experiment.

B. Plasma, serum and viral stock solution: prepare 10-200 μl 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 10 mg of virus-infected tissues to be ground with liquid nitrogen, and add 200 μl of PBS solution to the ground tissues.

Protocol A: Automatic Operation Process of Single Deep-Well Plate

Add the reagents listed below to each well:

Well	Reagent and Amount/Well	Operation process
A	Sample: 200 μl	Mix at 20~55°C for 10 min by vibrating and allow to settle for 2 min. The digested sample will release DNA/RNA to the Mag Beads. Transfer the beads to well B.
	Buffer MVL: 600 μl
	Mag Beads: 20 μl
	DS Carrier: 2 μl
	Proteinase K: 20 μl
B	Buffer MW: 600 μl	Wash the beads by vibrating for 2 min, then transfer the beads to well C.
C	Buffer MW: 600 μl	Wash the beads by vibrating for 2 min, dry the beads for 2 min, then transfer the beads to well D.
D	DEPC-Water: 200 μl	Elute by vibrating for 2 min, then transfer the beads back to well C.

The nucleic acid in well D is purified and can be used in RT-PCR, NGS, and other experiments or stored under -20°C.

Protocol B: Automatic Operation Process of 96 Deep-Well Plate

Add the reagents listed below to each plate:

Plate	Reagent and amount/well	Operation process
A	Sample: 200 μl	Mix at 20~55°C for 10 min by vibrating and allow to settle for 2 min. The digested sample will release DNA/RNA to the Mag Beads. Transfer the beads to well B.
	Buffer MVL: 600 μl
	Mag Beads: 20 μl
	DS Carrier: 2 μl
	Proteinase K: 20 μl
B	Buffer MW: 600 μl	Wash the beads by vibrating for 2 min, then transfer the beads to plate C.
C	Buffer MW: 600 μl	Wash the beads by vibrating for 2 min, dry the beads for 2 min, then transfer the beads to plate D.
D	DEPC-Water: 200 μl	Elute by vibrating for 2 min, then transfer the beads back to plate C.

The nucleic acid in plate D is purified and can be used in RT-PCR, NGS and other experiments or stored under -20°C.

Protocol C: Manual Operation Process

1. Prepare 200 μl sample in a 1.5 ml nuclease-free microcentrifuge tube. If the initial amount is less than 200 μl, use PBS solution to make it up to 200 μl.

2. Add the reagents in order: 600 μl Buffer MVL, 20 μl Mag Beads, 20 μl Proteinase K and 2 μl DS Carrier. Vortex vigorously for 15 sec to mix well. Incubate at room temperature for 10 min and mix by turning upside down twice during the process.

3. Centrifuge briefly. Place the tube on the magnetic rack and let it stand for 1 min. Remove the supernatant with a pipette.

4. Washing 1: Take the tube off the magnetic rack. Add 600 μl of Buffer MW. Vortex for 15 sec, then centrifuge briefly. Put the tube back on the magnetic rack and let stand for 1 min. Carefully discard all solutions.

5. Washing 2: Take the tube off the magnetic rack. Add 600 μl of Buffer MW. Vortex for 15 sec, then centrifuge briefly. Put the tube back on the magnetic rack, and let stand for 1 min. Carefully discard all solutions.

6. Air-dry at room temperature for 3 ~ 5 min until the surface of the magnetic beads does not reflect light.

Note: To ensure the purity of nucleic acid, no residual Buffer MW is allowed. The excessive drying (cracking) of the beads can affect the final yield.

7. Add 50 μl of DEPC-Water, vortex for 15 sec, and let stand for 3-5 min, during which gently oscillate 2 times to accelerate nucleic acid dissolution.

8. Put the tube back on the magnetic rack and let stand for 1 min. Pipette the supernatant to a new 1.5 ml

nuclease-free microcentrifuge tube. The obtained DNA/RNA can be directly used for subsequent detection, or be stored at -30 to -15°C for short-term storage or at -70°C for long-term storage.

Example of Application Allsheng Auto-Pure32A Automated Nucleic Acid Extractor

Step	Well	Name	Mixing Time	Magnetic Absorption Time	Waiting Time	Volume	Mixing Speed	Temperature
1	1	Lysis	10 min	0	2 min	840	8	OFF
2	1	Magnetic Absorption	0	90 sec	0	840	8	OFF
3	2	Wash 1	2 min	60 sec	2 min	600	8	OFF
4	3	Wash 2	2 min	60 sec	0	600	8	OFF
5	4	Elution	3 min	60 sec	0	50	8	OFF
6	1	Drop	0.5 min	0	0	840	8	OFF

This procedure can quickly and efficiently extract viral RNA from nasal swabs.

Product Number: GV4002

Ghana