u
Dispense Rate: 2000
4
Select Pump.
5
Inspect the flow cell for bubbles passing through the lanes and leaks near the manifolds.
6
If excessive bubbles are present, do as follows.
a
Check the gaskets for obstructions.
b
Reduce the aspirate rate to 100.
c
Pump another 125 µl of water to the flow cell.
d
If problems persist, remove the flow cell, repeat the cleaning steps, and reload the flow cell.
Position Tubing and Start Prime
1
Remove the 8 waste tubes for the appropriate flow cell from the waste container.
Figure 7
Position Tubing
A
Flow cell waste tubes for reagent positions 1–8
B
Condensation pump tubing
C
Paired-end priming pump tubing
2
Place each waste tube into a separate empty 15 ml tube.
Waste is collected and measured when priming is complete.
3
Select Start Prime. Monitor priming progress from the Prime screen.
4
When priming is complete, measure the waste and confirm that the volume in each tube is 1.75 ml for a
total of 14 ml.
The total is calculated as follows:
u
250 µl for each SBS position except position 2 (250 x 7 = 1.75 ml)
u
1.75 ml for each lane (1.75 x 8 = 14 ml)
5
Return the waste tubes to the waste container.
6
Select Next.
Load the Sequencing Flow Cell
Loading the flow cell for sequencing includes removing the priming flow cell, cleaning the flow cell holder,
loading the clustered flow cell, and confirming proper flow.
Document # 15011190 v03
For Research Use Only. Not for use in diagnostic procedures.
20
HiSeq 2000 System Guide