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Template

GTL Plate

EcoR1 MID plate

Pool Plate

Library

Norm to

Sequencer

WCR31

1

1+2

1

10

Next

WCR32

2

1+2

1

10

Next

WCR33

3

3

1

10

Next

WCR34

4

4+5

2

30

Next

WCR36

5

4+5

2

30

Next

WCR37

6

6+7

2

30

Next

WCR38

7

6+7

3

30

Next

WCR39

8

8+9

3

30

Next

WCR40

1

8+9

3

30

Next

WCR41

2

10+11

4

10

Next

WCR45

3

10+11

4

10

Next

WCR46

4

12

4

10

Next

WCR47

5

13+14

6

30

Nov

WCR48

6

13+14

6

30

Nov

WCR49

7

15+16

5

40

Nov

WCR50

8

15+16

5

40

Nov

WCR51

1

17+18

5

40

Nov

WCR52

2

19+20

6

30

Nov

WCR53

3

17+18

5

40

Nov

WCR54

4

19+20

6

30

Nov

WCR43

5

7

30

Nov

WCR44

6

7

30

Nov

WCR55

7

7

30

Nov

WCR56

8

7

30

Nov

WCR57

1

8

30

Next

WCR58

2

8

30

Next

WCR59

3

8

30

Next

WCR28

4

9

10*

Next

WCR35

5

9

10*

Next

WCR42

6

9

10*

Next

Label reaction plates with MID plate used.

...

Reagent

ul/rxn

rxns

ul needed (x1.4)

H2O

9.52

350

30941904

1332.8

5x iProof buffer

4

350

1300

800560

10 mM dNTPs

0.4

350

130

8056

50 mM MgCl2

0.4

350

130

8056

5 uM Illumina Primers

1.33

350

432266

186.2

iProof TAQ

0.2

350

65

4028

DMSO

0.15

350

49

3021

total

16

350

5200

32002240

Add 16 ul of PCR1 MM to each well of four new hard shell PCR plates with 8 channel

...

Reagent

ul/rxn

rxns

ul needed (x 1.6)

5x Iproof buffer

0.425

360

153

13565.15

10 mM dNTPs

0.4

360

144

12761.26

Primers

1.33

360

479423

204.8

Total

2.155

360

776

331.87

Then continue with the last four unlisted steps for GBS1 from above:

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