If the adjacent run is completing an imaging step, the software pauses the adjacent flow cell before you
can load and prime reagents.
2
After loading the sequencing flow cell for the new run, close the compartment door.
3
Select Start to start the new sequencing run.
Split SBS Kits
To perform a paired-end run that is shorter than 125 cycles per read, split a 250-cycle HiSeq v4 kit into 2 sets
of reagents. One set provides reagents for a 126-cycle run with dual indexing and 1 prime. For a run that is
shorter than 67 cycles, use a 50-cycle kit.
Splitting a 200-cycle TruSeq v3 kit provides enough reagents to perform up to 101 cycles with 1 prime. For a
run that is shorter than 101 cycles, use a 50-cycle kit.
Split a 250-Cycle Kit
1
Thaw reagents from the HiSeq v4 SBS kit (250 cycles). See
.
2
Label 7 250 ml bottles with the position number and reagent name, as follows.
u
#1 IRM
u
#3 USM
u
#4 SB1
u
#5 SB2
u
#6 SB2
u
#7 CRM
u
#8 SB3
3
Add the following volume of each reagent into the appropriate 250 ml bottle to result in a second set of
reagents. Prepare CRM last and then replace your gloves.
Number
Reagent Name
Volume
1
IRM
72 ml
3
USM
82 ml
4
SB1
100 ml
5
SB2 (for position #5)
62 ml
6
SB2 (for position #6)
62 ml
7
CRM
72 ml
8
SB3
110 ml
4
Use both sets of reagents for 2 concurrent runs or store 1 set at the following temperatures:
u
Store IRM, USM, and CRM at -25°C to -15°C.
u
Store SB1, SB2, and SB3 at 15°C to 30°C.
Split a 200-Cycle Kit
1
Thaw reagents from the TruSeq v3 SBS kit (200 cycles). See
2
Label 7 250 ml bottles with the position number and reagent name, as follows.
u
#1 ICB
u
#3 SRE
u
#4 SB1
Document # 15011190 v03
For Research Use Only. Not for use in diagnostic procedures.
53
HiSeq 2000 System Guide