Product Information(Virus DNA/RNA Extraction Kit)

Kit composition	Cat # HT-ZF	Storage
1. Adsorption column	50	2-30℃
2. Collect tubes	50	2-30℃
3. The proteinase K	1.1mL	2-8℃
4. Lysate Solution	15 mL	2-30℃
5. Wash Solution 1	21 mL	2-30℃
6. Wash Solution 2	7mL	2-30℃
7. Eluent	5mL	2-30℃

Note: Before you use, make sure Wash Solution 1 has been added 14mL absolute ethanol, and Wash Solution 2 has been added 28mL absolute ethanol.

Required Materials Not Provided With the Kit

• Anhydrous ethanol, 1.5 mL sterile EP tube, normal saline or PBS (pH 7.4) buffer, etc.
• Vortex mixer.
• 20, 100, 200 and 1000 mL finnpipettes.

Operation Procedure

Sample Preparation

1. Blood samples: Samples are centrifuged at 8000 rpm. for 2 minutes to take the upper serum which was placed in the sterilized centrifuge tube for reserve.
2. Tissue samples: Collect 0.1 g of diseased tissue samples from spleen, liver, lymph nodes and tonsils, fully homogenate or grind in tissue homogenizer, add 1 mL PBS (PH 7.4) buffer, centrifuged at 8000 rpm for 2 min, take supernatant in the sterilized centrifuge tube for reserve.
3. Oropharyngeal / nasal fluid samples: Samples are centrifuged at 8000 rpm. for 2 minutes, take supernatant in the sterilized centrifuge tube for reserve.
4. Feed: Put the appropriate amount of ground feed (about 10g) into 50 mL EP tubes containing 10 mL PBS (PH 7.4) buffer, shake and mix, centrifuge at 8000 rpm. for 2 minutes, take the supernatant in the sterilized centrifuge tube for reserve.
5. Environmental samples: Shake and mix, centrifuge at 8000 rpm. for 2 minutes, take the supernatant in the sterilized centrifuge tube for reserve.

Nucleic acid extraction

Amount of reagent required for one reaction is shown in the table below.

Note: To ensure the elution efficiency, the eluate should be added to the central position of the centrifugal column.

1. Add 250 mL lysate and 20 mL protease K into a new 1. 5 mL centrifuge tube.
2. Add 200 mL samples to the centrifuge tube, reverse mix for 10 times, and place lysis samples at room temperature for 5 mins. (if the room temperature is low, lysis should be replied to 25℃)
3. Add 250 mL anhydrous ethanol and mix in reverse for 10 seconds; if the sample is cloudy, centrifuge at 12000 rpm for 1 minute and remove the supernatant to avoid blocking the column;
4. Transfer the supernatant into an adsorption column with a collection tube and be centrifuged at 12000 rpm for 1 minute.
5. Discard the collection tube liquid, add 600 mL of Wash Solution1 to the adsorption column (please check whether 14 mL of absolute ethanol has been added), and centrifuge at 12,000 rpm for 1 minute;
6. Discard the collection tube liquid, add 600 mL of Wash Solution 2 to the adsorption column (please check whether 28 mL of absolute ethanol has been added), and centrifuge at 12,000 rpm for 1 minute;
7. Collection tube liquid was discarded and centrifuged at 12,000 rpm for 2 min to remove residual Wash Solution thoroughly.
8. The adsorption column was transferred into a new nuclease-free 1.5 mL EP tube, dropped 50 mL eluate to the center of the column, let for 1 minute, centrifuged at 12000 rpm for 1 minute. The liquid in the EP tube is nucleic acid. The nucleic acid is used immediately or stored -20℃.