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...

  •  Pull Plates Dibner_1-95_Norm, Dibner_96-190_Norm, MP101, and MP102 and allow to Thaw
  •  Tap plates to drop liquid, Spin to drop liquid
  •  SpeedVac to dry samples
  •  Resuspend Dibner_1-95_Norm and Dibner_96-190_Norm with 12 ul of ultra pure water
  •  Resuspend MP101 and MP102 with 25 ul ultra pure water
  •  Transfer the below samples to a new plate

plate_name

plate_position

sample_name

read_number

row

column

Dibner_1-95

A1

cp3_nb2_aug_19_19

2263

A

1

Dibner_1-95

B1

cp1_sf4_oct_18_19

1807

B

1

Dibner_1-95

D1

cp3_nb2_jun_24_19

7372

D

1

Dibner_1-95

G1

cp1_nb2_sep_23_19

512

G

1

Dibner_1-95

H1

cp1_nb2_may_22_19

1012

H

1

Dibner_1-95

A2

cp1_nb2_jul_23_19

12446

A

2

Dibner_1-95

B2

cp1_sf1_jun_20_19

1338

B

2

Dibner_1-95

C2

cp1_sf2_sep_23_19

1610

C

2

Dibner_1-95

D2

cp1_uf7_jul_23_19

10130

D

2

Dibner_1-95

E2

cp3_uf1_aug_19_19

2841

E

2

Dibner_1-95

F2

cp1_nb1_aug_19_19

2251

F

2

Dibner_1-95

G2

cp1_uf6_may_22_19

6034

G

2

Dibner_1-95

H2

cp1_uf3_apr_08_19

3132

H

2

Dibner_1-95

B3

cp1_uf6_sep_23_19

9789

B

3

Dibner_1-95

A4

cp1_uf4_apr_08_19

1701

A

4

Dibner_1-95

B4

cp1_uf5_apr_15_19

3837

B

4

Dibner_1-95

D4

1B_nana_nov_22_19

657

D

4

Dibner_1-95

G4

cp1_uf6_aug_19_19

742

G

4

Dibner_1-95

A5

cp2_sf3_aug_20_19

658

A

5

Dibner_1-95

B5

cp3_nb1_jun_24_19

19290

B

5

Dibner_1-95

C5

cp3_uf2_jun_24_19

6000

C

5

Dibner_1-95

A6

cp2_sf3_jun_21_19

9145

A

6

Dibner_1-95

H6

6B_nana_nov_22_19

141

H

6

Dibner_1-95

A7

cp1_nb4_sep_23_19

697

A

7

Dibner_1-95

E7

cp1_nb1_jun_20_19

3107

E

7

Dibner_1-95

B8

cp1_sf3_jun_20_19

3451

B

8

Dibner_1-95

B8 B7

cp1_sf4_jun_20_19

1626

B

8

Dibner_1-95

C8

cp1_uf5_jul_23_19

10685

C

8

Dibner_1-95

G8

cp1_nb2_mar_15_19

15710

G

8

Dibner_1-95

D9

cp1_sf3_aug_19_19

7424

D

9

Dibner_1-95

H9

5D_nana_nov_22_19

3085

H

9

Dibner_1-95

B10

cp1_nb2_aug_19_19

17950

B

10

Dibner_1-95

H10

cp2_nb2_jun_21_19

13715

H

10

Dibner_1-95

E11

cp1_sf5_may_22_19

2134

E

11

Dibner_1-95

C12

cp1_sf4_jul_23_19

1037

C

12

Dibner_1-95

H12

blank

21

H

12

Dibner_96-190

E3

cp1_nb5_apr_15_19

14423

E

3

Dibner_96-190

C4

cp1_uf5_jun_20_19

5544

C

4

Dibner_96-190

B5

cp1_sf1_sep_23_19

1435

B

5

MP101

G1

MP101_G1

1793

G

1

MP101

H1

MP101_H1

50

H

1

MP101

H4

MP101_H4

212

H

4

MP101

A11

MP101_A11

10565

A

11

MP102

A1

MP102_A1

1525

A

1

MP102

E1

MP102_E1

6866

E

1

MP102

B2

MP102_B2

491

B

2

MP102

H2

MP102_H2

1195

H

2

MP102

B3

MP102_B3

740

B

3

MP102

H3

MP102_H3

5542

H

3

MP102

D5

MP102_D5

758

D

5

MP102

D8

MP102_D8

374

D

8

MP102

H9

MP102_H9

453

H

9

MP102

G12

MP102_G12

8669

G

12

Well Position

Sample

A01

cp3_nb2_aug_19_19

B01

cp1_sf4_oct_18_19

C01

cp3_nb2_jun_24_19

D01

cp1_nb2_sep_23_19

E01

cp1_nb2_may_22_19

F01

cp1_nb2_jul_23_19

G01

cp1_sf1_jun_20_19

H01

cp1_sf2_sep_23_19

A02

cp1_uf7_jul_23_19

B02

cp3_uf1_aug_19_19

C02

cp1_nb1_aug_19_19

D02

cp1_uf6_may_22_19

E02

cp1_uf3_apr_08_19

F02

cp1_uf6_sep_23_19

G02

cp1_uf4_apr_08_19

H02

cp1_uf5_apr_15_19

A03

1B_nana_nov_22_19

B03

cp1_uf6_aug_19_19

C03

cp2_sf3_aug_20_19

D03

cp3_nb1_jun_24_19

E03

cp3_uf2_jun_24_19

F03

cp2_sf3_jun_21_19

G03

6B_nana_nov_22_19

H03

cp1_nb4_sep_23_19

A04

cp1_nb1_jun_20_19

B04

cp1_sf3_jun_20_19

C04

cp1_sf4_jun_20_19

D04

cp1_uf5_jul_23_19

E04

cp1_nb2_mar_15_19

F04

cp1_sf3_aug_19_19

G04

5D_nana_nov_22_19

H04

cp1_nb2_aug_19_19

A05

cp2_nb2_jun_21_19

B05

cp1_sf5_may_22_19

C05

cp1_sf4_jul_23_19

D05

blank

E05

cp1_nb5_apr_15_19

F05

cp1_uf5_jun_20_19

G05

cp1_sf1_sep_23_19

H05

MP101_G1

A06

MP101_H1

B06

MP101_H4

C06

MP101_A11

D06

MP102_A1

E06

MP102_E1

F06

MP102_B2

G06

MP102_H2

H06

MP102_B3

A07

MP102_H3

B07

MP102_D5

C07

MP102_D8

D07

MP102_H9

E07

MP102_G12

F07

Mock Community

G07

Mock Community

H07

Mock Community

...

  • Equilibrate Beads to room Temperature

  • Add 24 ul of MagBeads to each well and 15 ul of replicate to same well of replicate

  • Pipette mix up and down 10 times.

  • Incubate at RT for 5 minutes

  • Secure plate on magnet plate; incubate at RT for 5 minutes (until wells are clear)

  • Remove 65 ul from each well; keep tips to left or right depending on the column to avoid bead pellet.

  • Add 100 ul Fresh 80% EtOH to each well. Incubate 30 seconds. Remove 100 ul from each well

  • Add 100 ul Fresh 80% EtOH to each well. Incubate 30 seconds. Remove 100 ul from each well

  • Reaspirate from each well to assure maximum EtOH removal

  • Allow plate to air dry for 7 minutes.

  • Remove sample plate from magnet plate.

  • Add 40 ul TE; pipette mix 10+ times. Incubate 2 minutes at RT.

  • Place sample plate back on magnet for 5 minutes or until all wells are cleared.

  • Transfer 40 ul to labeled transparent plate (Plate1 PCR1 MIDPlate1 MIDPlate2)

  • Transfer 10 ul from transparent plate to “PCR2” plate with Mastermix already added.

  • Label Plate1 PCR2 MIDPlate1 MIDPlate2

  • Seal “PCR2”s with bubble strips and run on thermocycler 35GSAF2 program

...

Temp C

...

Cycles

...

Temp C

Cycles

Time

95*

1X

3:00

98

19X

0:30

55*

19X

0:30

72

19X

0:30

72

1X

5:00

4

1X

0:00

...