NOTE
For partial kits, the software counts down the number of cycles entered. When the cycles are low, the
software prompts for fresh reagents.
4
Select Prime SBS Reagents to prime reagents.
5
Select Next.
Review Screen
1
Review run parameters from the Review screen.
2
Select Next to proceed or Back to change parameters.
Load and Prime Reagents
After entering run parameters, load SBS, indexing, and paired-end reagents for the run, and then prime
reagents through the fluidics system. The software guides you through these steps in a series of screens on
the Pre-Run Setup tab.
Load SBS Reagents
1
Replace the cap on each reagent bottle with a funnel cap.
CAUTION
Handle the bottle of CMR last, after you have loaded all other reagents, to prevent cross-contamination.
Always replace your gloves after handling CMR.
2
Open the reagent compartment door.
3
Raise the sippers for the SBS reagent rack as follows.
a
Pull the sipper handle towards you and then raise it.
b
Release the handle into the slot on the top end of the groove. Make sure that the handle rests
securely in the slot.
4
Slide the reagent rack out of the reagent compartment.
5
Place each reagent bottle onto the rack in the associated numbered position. Make sure that the conical
end of the bottle rests in the indentation on the base of the rack.
Position
Reagent
Description
1
ICB
Incorporation Mix
2
PW1
25 ml of PW1 or laboratory-grade water
3
SRE
Scan Mix Reagent
4
SBS Buffer 1 (SB1)
High Salt Buffer
5
SBS Buffer 2 (SB2)
Incorporation Wash Buffer
6
SBS Buffer 2 (SB2)
Incorporation Wash Buffer
7
CMR
Cleavage Mix Reagent
8
SBS Buffer 3 (SB3)
Cleavage Buffer
6
Slide the reagent rack into the reagent compartment. Align the rack with the raised guide on the floor of
the compartment.
Document # 15011190 v03
For Research Use Only. Not for use in diagnostic procedures.
33
HiSeq 2000 System Guide