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Shannon Harris

Gregg Randolph

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  • Add 5 ul 1 nM Pool to 85 ul “10 mM Tris 8.5” and 10 ul 50 pM PhiX

  • Remove iSeq 100 i1 Flow Cell from refrigerator 5’s crisper drawer and open white foil pack and allow to equilibrate to RT for 10-15 minutes.

  • Open “iSeq 100 i1 Reagent Cartridge v2”. Turn on iSeq100

  • Click on “Sequence”. Watch Video. Do what video tells you to do. Follow on screen instructions until run starts.

Pooling for CU sequencing:

Sample.ID

Reads

Reads2

Reads3

Plate.Position

Plate

order

A26

35

26

18

A9

2

65

A27

42

20

15

B9

2

66

A29

16

7

4

C9

2

68

A30

36

24

11

D9

2

69

A31

40

20

15

E9

2

70

A32

28

18

11

F9

2

71

A34

36

12

8

G9

2

73

A35

34

18

10

H9

2

74

B3

392

248

149

G11

2

91

L108

77

47

32

F4

1

30

L54

70

33

19

F7

1

54

L88

416

236

143

A4

1

25

LB1

104

51

34

A2

1

9

LB2

164

107

45

F3

1

22

LB3

70

35

20

C5

1

35

LB4

308

194

159

H6

1

48

LB5

310

169

129

E8

1

61

LB6

118

68

50

B10

1

74

LB7

374

219

161

G11

1

87

Since the test sequencing included all samples, we are adjusting pooling for samples that both replicates returned less than 500 reads each. These samples will be pooled at a volume of 20 ul while all others will be pooled at 2 ul.