MolPure™ Magnetic Swab Viral DNA/RNA Kit

Product Information

Product Name	Cat#	Size
	18300ES50	50 T
MolPureTM Magnetic Swab Viral DNA/RNA Kit	18300ES60	100 T
	18300ES70	200 T

Product Description

MolPure^TM Magnetic Swab Viral DNA/RNA Kit can rapidly and efficiently isolate and purify high-purity viral DNA or RNA from swab and viral culture supernatant. The unique magnetic beads and carefully optimized buffer system are used to effectively capture the released nucleic acid. The extracted viral DNA or RNA is suitable for various downstream application experiments, such as PCR and fluorescence quantitative PCR, due to its high-purity, stable and reliable quality.

Package Information

Component Number	Components	Cat#/Size
		18300ES50 (50T)	18300ES60 (100T)	18300ES70 (200T)
18300-A	Lysis buffer*	10 mL	20 mL	40 mL
18300-B	Rinsing Solution*	15 mL	30 mL	60 mL
18300-C	Washing Solution*	12 mL	24 mL	48 mL
18300-D	Eluent	5 mL	10 mL	20 mL
18300-E	Magnetic bead suspension	1 mL	2 x 1 mL	4 x 1 mL
18300-F	Carrier RNA (from yeast)	310 µg	2 x 310 µg	4 x 310 µg
18300-G	RNase free H2O	0.5 mL	1 mL	2 mL

[Note]: 18300ES50 Lysis Buffer* requires 11 mL of Isopropanol. Rinsing Solution * requires 10.7 mL of Isopropanol. Washing Solution * requires 48 mL of Ethanol.

18300ES60 Lysis Buffer* requires 22 mL of Isopropanol. Rinsing Solution * requires 21.4 mL of Isopropanol. Washing Solution * requires 96 mL of Ethanol.

18300ES70 Lysis Buffer* requires 44 mL of Isopropanol. Rinsing Solution * requires 42.8 mL of Isopropanol. Washing Solution * requires 192 mL of Ethanol.

Methods of transportation and preservation

MolPure^TM Magnetic Swab Viral DNA/RNA Kit is stable at room temperature for 12 months.

Cautions

1. Check precipitation or turbidity for each solution when working in low environmental temperature to ensure the products are under normal working condition.

2. Add the required volume of Isopropanol or Ethanol as indicated on the label before use.

3. When used for the first time, 310 µL RNase-free H₂O (18300-G) is added to Carrier RNA (18300-F) to make the working solution (1 µg/µL). Aliquot and store at -20°C to avoid repeated freezing and thawing. Carrier RNA working solution is stable at -20°C for 12 months.

4. Avoid repeated freezing and thawing for frozen samples to ensure the quality of DNA and RNA in the samples.

5. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

Preparation before the experiment

Self-provided equipment and reagents: magnetic separation frame or automatic extraction instrument, water bath or metal bath, whirlpool oscillator, centrifuge, 1.5 mL centrifuge tube, etc.

Method of application

Step 1. Manual extraction method.

1.1 Processing method for nasal swab/pharyngeal swab sample: collect the sample and mix with normal saline or virus swab preservation solution by inverting or vortexing the tube. transfer 200 µL of liquid to a 1.5 mL nuclease-free centrifuge tube.

Note: If the sample is less than 200 µL, extra swab preservation solution or normal saline is added to bring the volume to 200 µL.

1.2 Add 400 µL Lysis Buffer* and 6 µL of Carrier RNA Solution, then shake well immediately.

Note: It is recommended to freshly prepare the lysis binding solution with Lysis Buffer and Carrier RNA Solution in one big tube based on the quantity of samples. Mix well and add 400 µL to each sample. The lysis binding solution is stable at 4°C for 1 hour.

1.3 Add 20 µL of magnetic bead suspension.

Note: Resuspend and mix well the magnetic beads before use and after adding to 4-5 samples.

1.4 Incubate at room temperature (15-25°C) for 10 minutes. Shake and mix every 2-3 minutes. Centrifuge briefly to get the solution down.

1.5 Place the centrifuge tube on the magnetic rack. After the magnetic beads are completely adsorbed, carefully remove the liquid.

1.6 Add 500 µL of Rinsing Solution* into the centrifuge tube, shake well and vortex for 1 min. Centrifuge briefly to get the solution down.

1.7 Put the centrifuge tube back on the magnetic rack. After the magnetic beads are completely absorbed, carefully remove the liquid.

1.8 Add 500 µL of Washing Solution* to the centrifuge tube. Shake well and vortex for 1 min. Centrifuge briefly to get the solution down.

1.9 Put the centrifuge tube back on the magnetic rack. After the magnetic beads are completely adsorbed, carefully remove the liquid.

1.10 Repeat steps 8 and 9 again.

1.11 Place the centrifuge tube on a magnetic stand, open the cap and dry at 56°C for 3-5 minutes to remove the residual ethanol.

Note: Be careful to ensure that the ethanol is completely evaporated, but the magnetic beads should not over-dried. Dry till the magnetic beads are just cracked.

1.12 Remove the centrifuge tube from the magnetic stand, add 50-100 µL of Eluent (50 µL recommended) into the centrifuge tube. Shake and mix evenly to disperse the magnetic beads and set at 56°C for 2 minutes. Centrifuge briefly to get the solution down.

1.13 Place the centrifuge tube back on the magnetic stand. After the magnetic beads are completely adsorbed, carefully transfer the liquid to a new nuclease-free centrifuge tube.

1.14 The solution can be stored at -20°C for short-term storage and at -80°C for long-term storage.

 

 

 

 

 

 

Step 2. The method of automatic extraction

Automatic instruments are used, taking Aosheng 32-channel nucleic acid extraction instrument as an example.

2.1 Add the corresponding reagents to the 96-well deep-well plate according to the following table (the samples should be equilibrated to room temperature):

The name of the column	Location	Reagent name	Dosage/hole
Sample processing column	Column 1/7	sample	200 µL
		Lysis buffer *	400 µL
		Carrier RNA solution	6 µL
Rinse	Column 2/8	Rinsing solution *	500 µL
Washing + magnetic bead column	Column 3/9	Washing solution * + magnetic bead suspension	500 µL + 20 µL
Washing column	Column 4/10	Washing solution *	500 µL
Elution column	Column 6/12	Eluent	70 µL

[Note]:

1) Fully resuspend the magnetic bead suspension before use and after adding to 4-5 samples.

2) It is recommended to freshly prepare the lysis binding solution with Lysis Buffer and Carrier RNA Solution in one big tube based on the quantity of samples. The lysis binding solution is stable at 4°C for 1 hour.

2.2 Correctly place the 96-well plate and the magnet sleeve according to the position of the extractor.

2.3 Follow the procedure below to perform automated extraction experiments.

2.4 After the automated procedure, transfer the eluates from columns 6 and 12 to a clean nuclease-free centrifuge tube. The solution can be stored at -20°C for short-term storage and at -80°C for long-term storage.

 

Extraction procedure for the Orsheng 32-channel nucleic acid extraction instrument

Step	Hole site	Mix time (min)	Mag- netic suction time (sec)	Waiting time (min)	volume (µL)	Mixing speed (1-10)	Tempe- rature (°C )	Mixed position (0-100%)	Mixed range (1-100%)	Mag- netic suction position (0-100%)	Mag- netic suction speed (1-10)
Take the magnetic beads	3	0.3	70	0	500	7	/	0	80	0	1
Split binding	1	3	60	0	600	5	/	0	80	0	1
Washing 1	2	1	40	0	500	7	/	0	80	0	1
Washing 2	3	1	40	0	500	5	/	0	80	0	1
Washing 3	4	1	40	1.5	500	5	/	0	80	0	1
Elution	6	1.5	70	0	70	6	56	0	80	0	1
Abandon magnetic beads	3	0.3	0	0	500	7	/	0	80	0	1

[Note]: Select "Synchronous heating action" in "Options", and select "Magnetic attraction in 9 segments" for the magnetic attraction method.