Exosome Sample Delivery

Note: No RNA protectant (eg: Trizol, RNA later) should be added to the samples prior to the separation of exosome. 

Tips：

1. Serum Samples
1) After collecting blood, gently pour the whole blood into a clean EP tube or centrifuge tube;
2) Placing the whold blood at 4°C for 3-4 hours for the blood clots to separate (it can also be placed in a 4°C refrigerator overnight);
3) Centrifugation at 5000 rpm and 4°C for 10min till the light yellow serum to reveal; after the supernatant is removed, have another centrifugation at 3000 rpm and 4°C for 10min to ensure the best serum quality;
4) Separate the serum and store it at -80°C before delivery.
Note: The sample delivery volume is suggested to be more than 4mL.
2. Plasma samples (Not to be anticoagulated with heparin)
1) Drain whole blood with a blood collection needle and an anticoagulation tube (including EDTA) and mix;
2) Placing the whold blood at 4°C for 3-4 hours, centrifuge at 5000 rpm and 4°C for 5min; take the supernatant fraction as plasma;
3) The plasma can be separately stored at -80°C before delivery.
Note: The sample delivery volume is suggested to be more than 4mL.
3. Serum Samples with exosome removed (prepared or purchased);
1) When the cell confluence rate of the culture dish reaches 80%~90%;
2) Aspirate the original medium and add appropriate trypsin (to cover the cells) for digestion;
3) After the cells are rounded, add an equal volume of serum-containing medium to terminate digestion;
4) Gently blow the cells with a pipette to suspend the cells;
5) Centrifuge for 5 minutes at 1100 rpm;
6) Aspirate the supernatant and wash the cells with medium removed from exosome serum;
7) Centrifuge for 5 minutes at 1100 rpm;
8) Aspirate the supernatant and add the medium without exosome serum to suspend the cells;
9) Transfer the cells to different culture dishes according to the cell types (the cell confluence rate is 60%~80% when the supernatant is collected) for further culture;
10) Collect the cell supernatant after 48h~72h culture
11) Centrifuge at 2000 rpm for 10 min, followed by another centrifuge at 10,000 rpm for 30 min to remove cells or cell debris, and then collect the supernatant
Note: The sample delivery volume is suggested to be more than 20mL.
4. Urine Sample
1) Collect urine into a 15mL centrifuge tube;
2) Centrifuge at 2500~3000rpm for 15min, remove the cells and take the supernatant;
3) Store the urine sample at -80°C before delivery.

Note: The sample delivery volume is suggested to be more than 10mL.
Serum, plasma and exosome belong to the special sample types with higher risk during extraction. RiboBio does not guarantee the success rate of extraction.