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1. Remove the desired number of 6F (SteriVacc) bags from their sterile pouches and place near the sterile docking device.
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2. Remove the transfer set TS91710 (“TS”) from the sterile pouch and close the clamps. The syringe contains air.
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3. Observe the “OFF” printed on the two way stopcock handle to become familiar with the on and off positions. Note the word “OFF” points to the tube that is closed off.
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4. Place the TS close to the sterile docking device.
5. Hang the source reservoir nearby.
6. Connect the inlet tubing on the TS two-way stopcock to the source reservoir. The inlet tubing is the tube that extends straight down from the syringe.
7. The outlet tube extends perpendicular to the syringe. Connect the outlet tubing to the SteriVacc container.
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8. Open the clamp on the source reservoir line.
9. Turn the stopcock to open the inlet to the Source Reservoir.
10. Push the syringe plunger in to expel air into the source reservoir.
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11. Hold the syringe with the tip facing down.
12. Draw the syringe plunger back (which pulls some of the air back again) until the desired volume from the source reservoir (not including air) is in the syringe.
13. Close the clamp on the tubing from the source reservoir.
14. Open the stopcock to the direction of the SteriVacc container.
15. Open the clamp to the SteriVacc container.
16. Hold the syringe with the tip facing down;
17. Push the aliquot into the SteriVacc container, clearing the tubing with the air from the syringe. It is necessary to deliver enough air into the SteriVacc container to allow clearing of the tubing.
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18. Hang or hold the SteriVacc container with its tubing up, to allow the air bubble to be drawn back with the syringe.
19. Leave a small amount of air at the point where seal will be made.
20. Close the tubing clamp to the container.
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21. Place the SteriVacc container into the bar sealer and seal the container with a first seal nearest to the tubing. Make a second seal slightly nearer to the container, and finally a third seal, again slightly nearer to the container.
If preferred, the PVC tubing may be sealed as near the bottom of the bag as possible and left attached in lieu of sealing and cutting off the bottom of the bag . Care should be used, as the PVC tubing will be brittle at temperatures near 42° C.
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22. Cut the tubing from the container between the first and second seals made.
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23. Tie a tag to the container using the hole in the side. Most sticky labels will eventually fall off these bags because the bags are nonstick. A sticky label should not be used under liquid nitrogen conditions.
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24. Sterile dock a new SteriVacc container to the tubing that remains from the first sterile connection. This process will disconnect what remains of the first SteriVacc bag.
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25. Repeat the steps listed above to fill and seal additional SteriVac containers.
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