1. Sample handling
1) Tissue treatment: cut small pieces of tissue and add appropriate PBS or 0.9% normal saline,
grind the tissue thoroughly with electric homogenizer, centrifugate and take the supernatant
for the second step of lysis.
2) Cotton swab treatment: transfer the swab wiped in the throat, nose and other parts to a 2ml
centrifuge tube, add a proper amount of PBS (200μl PBS soaking overnight is better), take the
supernatant for the second step of lysis.
3) Cells: direct lysis method: the second step of lysis is carried out immediately.
2. Take 200μl of the sample after the first step of treatment, add 200μl of buffer GTC, vortex
shaking for 30s, 56 ºC water bath for 10min.
3. Add 250μl of national drug grade isopropanol and let stand for 5min at room temperature.
Transfer all the solutions to the adsorption column, centrifugate at 12000rpm for 1min,
discard the waste liquid in the collection tube, and put the adsorption column into the
collection tube again.
4. Add 600μl wash buffer to the adsorption column, centrifugate at 12000rpm for 1min,
discard the waste liquid in the collection tube, and then put the adsorption column into the
collection tube again.
5. Repeat step 4.
6.Put the adsorption column in the collection pipe and centrifugate at 12000rpm for 2min.
Note: the steps should not be omitted to ensure that the residual cleaning solution of the
adsorption column is removed.
Put the adsorption column in a clean 1.5ml centrifuge tube, open the cover of the adsorption
column, leave it at room temperature for 2min, add 30-50μl eluent buffer the adsorption
column, leave it at room temperature for 2-5min, and centrifugate it at 12000rpm for 2min to
obtain viral nucleic acid. Short term storage at - 80 ºC