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Received New DNAs 9-6-2022

Digests started 9-12-2022

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Add 1 ul of EcoR1 Adaptors with 8-channel and Track which template plate gets which MID plate.

Template

EcoR1 MID plate

JJ_AW_1 (1SMIN11BUN2 (1PJD2)

EcoR1 MID plate 1

JJ_AW_2 (1SMIN2)

EcoR1 MID plate 2

PJDB_JJF (1BUN1)

EcoR1 MID plate 3

4

Label reaction plates with MID plate used.

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Create working pools. Load Pool A1 into columns 1-6 and A2 into columns 7-12 of the pooling plate. 

Pool

Plate1 Col 1-6

Plate1 Col 7-122

Plate2 Plate1 Col 1-63

Plate2 Col 7-12

1Smin Pool A1

4

 

Pool

Plate1 Col 1-6

Plate1 Col 12-7

Plate2 Col 1-6

Plate2 Col 12-7

1Smin Pool A2

1

2

3

4

5

6

7

8

9

10

11

12

A

PLT1_Col1

PLT1_Col7

PLT2_Col1

PLT2_Col7

PLT1_Col2

PLT1_Col8

PLT2_Col2

PLT2_Col8

PLT1_Col3

PLT1_Col9

PLT2_Col3

PLT2_Col9

PLT1_Col4

PLT1_Col10

PLT2_Col4

PLT2_Col10

PLT1_Col5

PLT1_Col11

PLT2_Col5

PLT2_Col11

PLT1_Col6

PLT1_Col12

PLT2_Col6

PLT2_Col12

PLT1_Col1

PLT1_Col12

PLT2_Col1

PLT2_Col12

PLT1_Col2

PLT1_Col11

PLT2_Col2

PLT2_Col11

PLT1_Col3

PLT1_Col10

PLT2_Col3

PLT2_Col10

PLT1_Col4

PLT1_Col9

PLT2_Col4

PLT2_Col9

PLT1_Col5

PLT1_Col8

PLT2_Col5

PLT2_Col8

PLT1_Col6

PLT1_Col7

PLT2_Col6

PLT2_Col7

B

C

D

E

F

G

H

Pool

Plate3 Rows A-B

Plate3 Rows C-D

Plate3 Rows E-F

Plate3 Rows G-H

PJDA

 

Pool

Plate3 Rows A-B

Plate3 Rows D-C

Plate3 Rows E-F

Plate3 Rows H-G

PJDB

Pool

Plate3

Plate3

Plate3

Plate3

Mix1

A7, C7, E7, G7

B7, D7, F7, H7

Pool

Plate3

Plate3

Plate3

Plate3

Mix2

A7-D7

E7-H7

1

2

3

4

5

6

7

8

9

10

11

12

A

PLT3_Row A, Row C, Row E, Row G

PLT3_Row B, Row D, Row F, Row H

PLT3_Row A, Row D, Row E, Row H

PLT3_Row B, Row C, Row F, Row G

PLT3_A7, C7, E7, G7

B

PLT3_B7, D7, F7, H7

C

PLT3_A7, B7, C7, D7

D

PLT3_E7, F7, G7, H7

E

F

G

H

2 (flipped)

Plate1 Col 3

Plate2 Col 4(flipped)

 

PCR1:

Make MM1 in a 1.5mL tube:

Reagent

ul/rxn

rxns (x1.3)

ul needed

H2O

9.52

14816

1837.4238

5x iProof buffer

4

14816

772100

10 mM dNTPs

0.4

14816

77.210

50 mM MgCl2

0.4

14816

77.210

5 uM Illumina Primers

1.33

14816

25633.725

iProof TAQ

0.2

14816

38.65

DMSO

0.15

14816

293.75

total

16

14816

3088400

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

...

148

Reagent

ul/rxn

rxns (x1.6)

ul needed

5x Iproof buffer

0.425

16

10010.72

10 mM dNTPs

0.4

14816

949.86

Primers

1.33

14816

31531.29

Total

2.155

14816

51051.7

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

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