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C

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

0.002 pM Std

...

0.002 pM Std

...

0.002 pM Std

...

D

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

0.02 pM Std

...

0.02 pM Std

...

0.02 pM Std

...

E

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

0.2 pM Std

...

0.2 pM Std

...

0.2 pM Std

...

F

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

2 pM Std

...

2 pM Std

...

2 pM Std

...

G

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

20 pM Std

...

20 pM Std

...

20 pM Std

...

H

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

 

...

 

...

 

Results:

Average results for the following plates (column 3):

5CM1_16S: 6.03 nanomoles

5CM2_16S: 9.32 nanomoles

5CM3_16S: 3.52 nanomoles

5CM4_16S: 9.77 nanomoles

Results for HMax_PP_Final can be viewed in result report below and have been added to the HMax page here.

Full result report can be viewed below:

Pooling and Shipping:

The client, Chris MacGlover, is concerned about the nasal swabs not sequencing as occurred with his last attempt. We will send 4 nasal and one other sample to be fragment analyzed at CU:

5CM1 A4 (18-143N), 5CM1 H12 (SB33N), 5CM2 A4 ( 18-143T), 5CM3 H12 (51N), and 5CM1 A3 (excontrol)

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UWYO Plate Name 

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Prev Plate Name 

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Well

...

Sample Name

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5CM1

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Nasal 1A

...

A01

...

blank

...

5CM1

...

Nasal 1A

...

A02

...

blank

...

5CM1

...

Nasal 1A

...

A03

...

excontrol

...

5CM1

...

Nasal 1A

...

A04

...

18-143N

...

5CM1

...

Nasal 1A

...

A05

...

18-145N

...

5CM1

...

Nasal 1A

...

A06

...

18-150N

...

5CM1

...

Nasal 1A

...

A07

...

18-152N

...

5CM1

...

Nasal 1A

...

A08

...

18-157N

...

5CM1

...

Nasal 1A

...

A09

...

18-163N

...

5CM1

...

Nasal 1A

...

A10

...

18-168N

...

5CM1

...

Nasal 1A

...

A11

...

18-169N

...

5CM1

...

Nasal 1A

...

A12

...

18-171N

...

5CM1

...

Nasal 1A

...

B01

...

18-172N

...

5CM1

...

Nasal 1A

...

B02

...

18-173N

...

5CM1

...

Nasal 1A

...

B03

...

18-174N

...

5CM1

...

Nasal 1A

...

B04

...

18-175N

...

5CM1

...

Nasal 1A

...

B05

...

18-177N

...

5CM1

...

Nasal 1A

...

B06

...

18-178N

...

5CM1

...

Nasal 1A

...

B07

...

18-180N

...

5CM1

...

Nasal 1A

...

B08

...

18-181N

...

5CM1

...

Nasal 1A

...

B09

...

18-182N

...

5CM1

...

Nasal 1A

...

B10

...

18-183N

...

5CM1

...

Nasal 1A

...

B11

...

18-184N

...

5CM1

...

Nasal 1A

...

B12

...

18-185N

...

5CM1

...

Nasal 1A

...

C01

...

18-186N

...

5CM1

...

Nasal 1A

...

C02

...

18-187N

...

5CM1

...

Nasal 1A

...

C03

...

18-188N

...

5CM1

...

Nasal 1A

...

C04

...

18-189N

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5CM1

...

Nasal 1A

...

C05

...

18-190N

...

5CM1

...

Nasal 1A

...

C06

...

18-191N

...

5CM1

...

Nasal 1A

...

C07

...

19-001N

...

5CM1

...

Nasal 1A

...

C08

...

19-002N

...

5CM1

...

Nasal 1A

...

C09

...

19-003N

...

5CM1

...

Nasal 1A

...

C10

...

19-004N

...

5CM1

...

Nasal 1A

...

C11

...

19-005N

...

5CM1

...

Nasal 1A

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C12

...

19-006N

...

5CM1

...

Nasal 1A

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D01

...

19-007N

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5CM1

...

Nasal 1A

...

D02

...

19-008N

...

5CM1

...

Nasal 1A

...

D03

...

19-010N

...

5CM1

...

Nasal 1A

...

D04

...

19-011N

...

5CM1

...

Nasal 1A

...

D05

...

19-013N

...

5CM1

...

Nasal 1A

...

D06

...

19-015N

...

5CM1

...

Nasal 1A

...

D07

...

19-017N

...

5CM1

...

Nasal 1A

...

D08

...

19-018N

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5CM1

...

Nasal 1A

...

D09

...

19-019N

...

5CM1

...

Nasal 1A

...

D10

...

19-020N

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5CM1

...

Nasal 1A

...

D11

...

19-021N

...

5CM1

...

Nasal 1A

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D12

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19-023N

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5CM1

...

Nasal 1A

...

E01

...

19-025N

...

5CM1

...

Nasal 1A

...

E02

...

19-026N

...

5CM1

...

Nasal 1A

...

E03

...

19-027N

...

5CM1

...

Nasal 1A

...

E04

...

19-028N

...

5CM1

...

Nasal 1A

...

E05

...

19-029N

...

5CM1

...

Nasal 1A

...

E06

...

19-030N

...

5CM1

...

Nasal 1A

...

E07

...

19-031N

...

5CM1

...

Nasal 1A

...

E08

...

19-032N

...

5CM1

...

Nasal 1A

...

E09

...

19-033N

...

5CM1

...

Nasal 1A

...

E10

...

19-034N

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5CM1

...

Nasal 1A

...

E11

...

19-036-N

...

5CM1

...

Nasal 1A

...

E12

...

19-037N

...

5CM1

...

Nasal 1A

...

F01

...

19-038N

...

5CM1

...

Nasal 1A

...

F02

...

19-040N

...

5CM1

...

Nasal 1A

...

F03

...

19-041N

...

5CM1

...

Nasal 1A

...

F04

...

19-042N

...

5CM1

...

Nasal 1A

...

F05

...

19-043N

...

5CM1

...

Nasal 1A

...

F06

...

19-044N

...

5CM1

...

Nasal 1A

...

F07

...

19-047N

...

5CM1

...

Nasal 1A

...

F08

...

19-048N

...

5CM1

...

Nasal 1A

...

F09

...

19-049N

...

5CM1

...

Nasal 1A

...

F10

...

19-050N

...

5CM1

...

Nasal 1A

...

F11

...

19-051N

...

5CM1

...

Nasal 1A

...

F12

...

19-052N

...

5CM1

...

Nasal 1A

...

G01

...

19-053N

...

5CM1

...

Nasal 1A

...

G02

...

19-054N

...

5CM1

...

Nasal 1A

...

G03

...

19-055N

...

5CM1

...

Nasal 1A

...

G04

...

19-056N

...

5CM1

...

Nasal 1A

...

G05

...

19-058N

...

5CM1

...

Nasal 1A

...

G06

...

19-059N

...

5CM1

...

Nasal 1A

...

G07

...

19-060N

...

5CM1

...

Nasal 1A

...

G08

...

19-061N

...

5CM1

...

Nasal 1A

...

G09

...

19-062N

...

5CM1

...

Nasal 1A

...

G10

...

19-063N

...

5CM1

...

Nasal 1A

...

G11

...

19-064N

...

5CM1

...

Nasal 1A

...

G12

...

19-065N

...

5CM1

...

Nasal 1A

...

H01

...

C26N

...

5CM1

...

Nasal 1A

...

H02

...

C27N

...

5CM1

...

Nasal 1A

...

H03

...

C30N

...

5CM1

...

Nasal 1A

...

H04

...

C31N

...

5CM1

...

Nasal 1A

...

H05

...

C32N

...

5CM1

...

Nasal 1A

...

H06

...

C33N

...

5CM1

...

Nasal 1A

...

H07

...

C35 N

...

5CM1

...

Nasal 1A

...

H08

...

SB26N

...

5CM1

...

Nasal 1A

...

H09

...

SB27N

...

5CM1

...

Nasal 1A

...

H10

...

SB30N

...

5CM1

...

Nasal 1A

...

H11

...

SB31N

...

5CM1

...

Nasal 1A

...

H12

...

SB33N

...

5CM2

...

Tonsil 1B

...

A01

...

blank

...

5CM2

...

Tonsil 1B

...

A02

...

blank

...

5CM2

...

Tonsil 1B

...

A03

...

excontrol

...

5CM2

...

Tonsil 1B

...

A04

...

18-143T

...

5CM2

...

Tonsil 1B

...

A05

...

18-145T

...

5CM2

...

Tonsil 1B

...

A06

...

18-150T

...

5CM2

...

Tonsil 1B

...

A07

...

18-152T

...

5CM2

...

Tonsil 1B

...

A08

...

18-154T

...

5CM2

...

Tonsil 1B

...

A09

...

18-157T

...

5CM2

...

Tonsil 1B

...

A10

...

18-163T

...

5CM2

...

Tonsil 1B

...

A11

...

18-168T

...

5CM2

...

Tonsil 1B

...

A12

...

18-169T

...

5CM2

...

Tonsil 1B

...

B01

...

18-171T

...

5CM2

...

Tonsil 1B

...

B02

...

18-172T

...

5CM2

...

Tonsil 1B

...

B03

...

18-173T

...

5CM2

...

Tonsil 1B

...

B04

...

18-174T

...

5CM2

...

Tonsil 1B

...

B05

...

18-175T

...

5CM2

...

Tonsil 1B

...

B06

...

18-177T

...

5CM2

...

Tonsil 1B

...

B07

...

18-178T

...

5CM2

...

Tonsil 1B

...

B08

...

18-180T

...

5CM2

...

Tonsil 1B

...

B09

...

18-181T

...

5CM2

...

Tonsil 1B

...

B10

...

18-182T

...

5CM2

...

Tonsil 1B

...

B11

...

18-183T

...

5CM2

...

Tonsil 1B

...

B12

...

18-184T

...

5CM2

...

Tonsil 1B

...

C01

...

18-185T

...

5CM2

...

Tonsil 1B

...

C02

...

18-186T

...

5CM2

...

Tonsil 1B

...

C03

...

18-187T

...

5CM2

...

Tonsil 1B

...

C04

...

18-188T

...

5CM2

...

Tonsil 1B

...

C05

...

18-189T

...

5CM2

...

Tonsil 1B

...

C06

...

18-190T

...

5CM2

...

Tonsil 1B

...

C07

...

18-191T

...

5CM2

...

Tonsil 1B

...

C08

...

19-001T

...

5CM2

...

Tonsil 1B

...

C09

...

19-002T

...

5CM2

...

Tonsil 1B

...

C10

...

19-003T

...

5CM2

...

Tonsil 1B

...

C11

...

19-004T

...

5CM2

...

Tonsil 1B

...

C12

...

19-005T

...

5CM2

...

Tonsil 1B

...

D01

...

19-006T

...

5CM2

...

Tonsil 1B

...

D02

...

19-007T

...

5CM2

...

Tonsil 1B

...

D03

...

19-008T

...

5CM2

...

Tonsil 1B

...

D04

...

19-010T

...

5CM2

...

Tonsil 1B

...

D05

...

19-011T

...

5CM2

...

Tonsil 1B

...

D06

...

19-013T

...

5CM2

...

Tonsil 1B

...

D07

...

19-015T

...

5CM2

...

Tonsil 1B

...

D08

...

19-017T

...

5CM2

...

Tonsil 1B

...

D09

...

19-018T

...

5CM2

...

Tonsil 1B

...

D10

...

19-019T

...

5CM2

...

Tonsil 1B

...

D11

...

19-020T

...

5CM2

...

Tonsil 1B

...

D12

...

19-021T

...

5CM2

...

Tonsil 1B

...

E01

...

19-023T

...

5CM2

...

Tonsil 1B

...

E02

...

19-025T

...

5CM2

...

Tonsil 1B

...

E03

...

19-026T

...

5CM2

...

Tonsil 1B

...

E04

...

19-027T

...

5CM2

...

Tonsil 1B

...

E05

...

19-028T

...

5CM2

...

Tonsil 1B

...

E06

...

19-029T

...

5CM2

...

Tonsil 1B

...

E07

...

19-030T

...

5CM2

...

Tonsil 1B

...

E08

...

19-031T

...

5CM2

...

Tonsil 1B

...

E09

...

19-032T

...

5CM2

...

Tonsil 1B

...

E10

...

19-033T

...

5CM2

...

Tonsil 1B

...

E11

...

19-034T

...

5CM2

...

Tonsil 1B

...

E12

...

19-036T

...

5CM2

...

Tonsil 1B

...

F01

...

19-037T

...

5CM2

...

Tonsil 1B

...

F02

...

19-038T

...

5CM2

...

Tonsil 1B

...

F03

...

19-040T

...

5CM2

...

Tonsil 1B

...

F04

...

19-041T

...

5CM2

...

Tonsil 1B

...

F05

...

19-042T

...

5CM2

...

Tonsil 1B

...

F06

...

19-043T

...

5CM2

...

Tonsil 1B

...

F07

...

19-044T

...

5CM2

...

Tonsil 1B

...

F08

...

19-047T

...

5CM2

...

Tonsil 1B

...

F09

...

19-048T

...

5CM2

...

Tonsil 1B

...

F10

...

19-049T

...

5CM2

...

Tonsil 1B

...

F11

...

19-050T

...

5CM2

...

Tonsil 1B

...

F12

...

19-051T

...

5CM2

...

Tonsil 1B

...

G01

...

19-052T

...

5CM2

...

Tonsil 1B

...

G02

...

19-053T

...

5CM2

...

Tonsil 1B

...

G03

...

19-054T

...

5CM2

...

Tonsil 1B

...

G04

...

19-055T

...

5CM2

...

Tonsil 1B

...

G05

...

19-056T

...

5CM2

...

Tonsil 1B

...

G06

...

19-058T

...

5CM2

...

Tonsil 1B

...

G07

...

19-059T

...

5CM2

...

Tonsil 1B

...

G08

...

19-060T

...

5CM2

...

Tonsil 1B

...

G09

...

19-061T

...

5CM2

...

Tonsil 1B

...

G10

...

19-062T

...

5CM2

...

Tonsil 1B

...

G11

...

19-063T

...

5CM2

...

Tonsil 1B

...

G12

...

19-064T

...

5CM2

...

Tonsil 1B

...

H01

...

19-065T

...

5CM2

...

Tonsil 1B

...

H02

...

C26T

...

5CM2

...

Tonsil 1B

...

H03

...

C27T

...

5CM2

...

Tonsil 1B

...

H04

...

C30T

...

5CM2

...

Tonsil 1B

...

H05

...

C31T

...

5CM2

...

Tonsil 1B

...

H06

...

C32T

...

5CM2

...

Tonsil 1B

...

H07

...

C33T

...

5CM2

...

Tonsil 1B

...

H08

...

C35 T

...

5CM2

...

Tonsil 1B

...

H09

...

SB26T

...

5CM2

...

Tonsil 1B

...

H10

...

SB27T

...

5CM2

...

Tonsil 1B

...

H11

...

SB30T

...

5CM2

...

Tonsil 1B

...

H12

...

SB31T

...

5CM3

...

N - T 1C

...

A01

...

blank

...

5CM3

...

N - T 1C

...

A02

...

blank

...

5CM3

...

N - T 1C

...

A03

...

excontrol

...

5CM3

...

N - T 1C

...

A04

...

SB33T

...

5CM3

...

N - T 1C

...

A05

...

B12T

...

5CM3

...

N - T 1C

...

A06

...

B31T

...

5CM3

...

N - T 1C

...

A07

...

B54T

...

5CM3

...

N - T 1C

...

A08

...

B60T

...

5CM3

...

N - T 1C

...

A09

...

BK04T

...

5CM3

...

N - T 1C

...

A10

...

BK122T

...

5CM3

...

N - T 1C

...

A11

...

BK128T

...

5CM3

...

N - T 1C

...

A12

...

BK13T

...

5CM3

...

N - T 1C

...

B01

...

BK19T

...

5CM3

...

N - T 1C

...

B02

...

BK20T

...

5CM3

...

N - T 1C

...

B03

...

BK21T

...

5CM3

...

N - T 1C

...

B04

...

BK23T

...

5CM3

...

N - T 1C

...

B05

...

BK24T

...

5CM3

...

N - T 1C

...

B06

...

BK52T

...

5CM3

...

N - T 1C

...

B07

...

BK5T

...

5CM3

...

N - T 1C

...

B08

...

BK83T

...

5CM3

...

N - T 1C

...

B09

...

BK91T

...

5CM3

...

N - T 1C

...

B10

...

BK98T

...

5CM3

...

N - T 1C

...

B11

...

BLU165T

...

5CM3

...

N - T 1C

...

B12

...

BLU20T

...

5CM3

...

N - T 1C

...

C01

...

R02T

...

5CM3

...

N - T 1C

...

C02

...

R03T

...

5CM3

...

N - T 1C

...

C03

...

R11T

...

5CM3

...

N - T 1C

...

C04

...

R16T

...

5CM3

...

N - T 1C

...

C05

...

R18T

...

5CM3

...

N - T 1C

...

C06

...

R20T

...

5CM3

...

N - T 1C

...

C07

...

R21T

...

5CM3

...

N - T 1C

...

C08

...

Y14T

...

5CM3

...

N - T 1C

...

C09

...

Y46T

...

5CM3

...

N - T 1C

...

C10

...

Y49T

...

5CM3

...

N - T 1C

...

C11

...

Y51T

...

5CM3

...

N - T 1C

...

C12

...

Y52T

...

5CM3

...

N - T 1C

...

D01

...

Y53T

...

5CM3

...

N - T 1C

...

D02

...

Y63T

...

5CM3

...

N - T 1C

...

D03

...

Y66T

...

5CM3

...

N - T 1C

...

D04

...

Y69T

...

5CM3

...

N - T 1C

...

D05

...

Y81T

...

5CM3

...

N - T 1C

...

D06

...

Y84T

...

5CM3

...

N - T 1C

...

D07

...

Y87T

...

5CM3

...

N - T 1C

...

D08

...

YA1T

...

5CM3

...

N - T 1C

...

D09

...

YA3T

...

5CM3

...

N - T 1C

...

D10

...

FR-Y53AT

...

5CM3

...

N - T 1C

...

D11

...

FR-Y53T

...

5CM3

...

N - T 1C

...

D12

...

HG-W120T

...

5CM3

...

N - T 1C

...

E01

...

19-8349T

...

5CM3

...

N - T 1C

...

E02

...

19-9053T

...

5CM3

...

N - T 1C

...

E03

...

B12N

...

5CM3

...

N - T 1C

...

E04

...

B31N

...

5CM3

...

N - T 1C

...

E05

...

B54N

...

5CM3

...

N - T 1C

...

E06

...

B60N

...

5CM3

...

N - T 1C

...

E07

...

BK04N

...

5CM3

...

N - T 1C

...

E08

...

BK122N

...

5CM3

...

N - T 1C

...

E09

...

BK128N

...

5CM3

...

N - T 1C

...

E10

...

BK13N

...

5CM3

...

N - T 1C

...

E11

...

BK19N

...

5CM3

...

N - T 1C

...

E12

...

BK20N

...

5CM3

...

N - T 1C

...

F01

...

BK21N

...

5CM3

...

N - T 1C

...

F02

...

BK23N

...

5CM3

...

N - T 1C

...

F03

...

BK24N

...

5CM3

...

N - T 1C

...

F04

...

BK52N

...

5CM3

...

N - T 1C

...

F05

...

BK5N

...

5CM3

...

N - T 1C

...

F06

...

BK83N

...

5CM3

...

N - T 1C

...

F07

...

BK91N

...

5CM3

...

N - T 1C

...

F08

...

BK98N

...

5CM3

...

N - T 1C

...

F09

...

BLU165N

...

5CM3

...

N - T 1C

...

F10

...

BLU20N

...

5CM3

...

N - T 1C

...

F11

...

R02N

...

5CM3

...

N - T 1C

...

F12

...

R03N

...

5CM3

...

N - T 1C

...

G01

...

R11N

...

5CM3

...

N - T 1C

...

G02

...

R16N

...

5CM3

...

N - T 1C

...

G03

...

R18N

...

5CM3

...

N - T 1C

...

G04

...

R20N

...

5CM3

...

N - T 1C

...

G05

...

R21N

...

5CM3

...

N - T 1C

...

G06

...

Y14N

...

5CM3

...

N - T 1C

...

G07

...

Y46N

...

5CM3

...

N - T 1C

...

G08

...

Y49N

...

5CM3

...

N - T 1C

...

G09

...

Y51N

...

5CM3

...

N - T 1C

...

G10

...

Y52N

...

5CM3

...

N - T 1C

...

G11

...

Y53N

...

5CM3

...

N - T 1C

...

G12

...

Y63N

...

5CM3

...

N - T 1C

...

H01

...

Y66N

...

5CM3

...

N - T 1C

...

H02

...

Y69N

...

5CM3

...

N - T 1C

...

H03

...

Y81N

...

5CM3

...

N - T 1C

...

H04

...

Y84N

...

5CM3

...

N - T 1C

...

H05

...

Y87N

...

5CM3

...

N - T 1C

...

H06

...

YA1N

...

5CM3

...

N - T 1C

...

H07

...

YA3N

...

5CM3

...

N - T 1C

...

H08

...

47N

...

5CM3

...

N - T 1C

...

H09

...

48N

...

5CM3

...

N - T 1C

...

H10

...

49N

...

5CM3

...

N - T 1C

...

H11

...

50N

...

5CM3

...

N - T 1C

...

H12

...

51N

...

5CM4

...

Nasal 2D

...

A01

...

blank

...

5CM4

...

Nasal 2D

...

A02

...

blank

...

5CM4

...

Nasal 2D

...

A03

...

 

...

5CM4

...

Nasal 2D

...

A04

...

64N

...

5CM4

...

Nasal 2D

...

A05

...

19-10733N

...

5CM4

...

Nasal 2D

...

A06

...

19-10734N

...

5CM4

...

Nasal 2D

...

A07

...

 

...

5CM4

...

Nasal 2D

...

A08

...

 

...

5CM4

...

Nasal 2D

...

A09

...

 

...

5CM4

...

Nasal 2D

...

A10

...

 

...

5CM4

...

Nasal 2D

...

A11

...

 

...

5CM4

...

Nasal 2D

...

A12

...

 

...

5CM4

...

Nasal 2D

...

B01

...

52N

...

5CM4

...

Nasal 2D

...

B02

...

56N

...

5CM4

...

Nasal 2D

...

B03

...

60N

...

5CM4

...

Nasal 2D

...

B04

...

65N

...

5CM4

...

Nasal 2D

...

B05

...

Y53AN

...

5CM4

...

Nasal 2D

...

B06

...

19-8349N

...

5CM4

...

Nasal 2D

...

B07

...

 

...

5CM4

...

Nasal 2D

...

B08

...

 

...

5CM4

...

Nasal 2D

...

B09

...

 

...

5CM4

...

Nasal 2D

...

B10

...

 

...

5CM4

...

Nasal 2D

...

B11

...

 

...

5CM4

...

Nasal 2D

...

B12

...

 

...

5CM4

...

Nasal 2D

...

C01

...

53N

...

5CM4

...

Nasal 2D

...

C02

...

57N

...

5CM4

...

Nasal 2D

...

C03

...

61N

...

5CM4

...

Nasal 2D

...

C04

...

66N

...

5CM4

...

Nasal 2D

...

C05

...

Y53BN

...

5CM4

...

Nasal 2D

...

C06

...

19-9053N

...

5CM4

...

Nasal 2D

...

C07

...

 

...

5CM4

...

Nasal 2D

...

C08

...

 

...

5CM4

...

Nasal 2D

...

C09

...

 

...

5CM4

...

Nasal 2D

...

C10

...

 

...

5CM4

...

Nasal 2D

...

C11

...

 

...

5CM4

...

Nasal 2D

...

C12

...

 

...

5CM4

...

Nasal 2D

...

D01

...

54N

...

5CM4

...

Nasal 2D

...

D02

...

58N

...

5CM4

...

Nasal 2D

...

D03

...

62N

...

5CM4

...

Nasal 2D

...

D04

...

67N

...

5CM4

...

Nasal 2D

...

D05

...

HG-W120AN

...

5CM4

...

Nasal 2D

...

D06

...

19-09761N

...

5CM4

...

Nasal 2D

...

D07

...

 

...

5CM4

...

Nasal 2D

...

D08

...

 

...

5CM4

...

Nasal 2D

...

D09

...

 

...

5CM4

...

Nasal 2D

...

D10

...

 

...

5CM4

...

Nasal 2D

...

D11

...

 

...

5CM4

...

Nasal 2D

...

D12

...

 

...

5CM4

...

Nasal 2D

...

E01

...

55N

...

5CM4

...

Nasal 2D

...

E02

...

59N

...

5CM4

...

Nasal 2D

...

E03

...

63N

...

5CM4

...

Nasal 2D

...

E04

...

68N

...

5CM4

...

Nasal 2D

...

E05

...

...

5CM1 (16S only)

...

5CM2 (16S only)

...

5CM3 (16S only)

...

5CM4 (16S only)

MasterMix (make for 8 plates in 15mL conical tube)

...

ul/rxn

...

Reagent

...

# of rxns

...

ul needed (1.5X)

...

3

...

5X Kapa HiFi Buffer

...

768

...

3456

...

0.45

...

10M dNTPs

...

768

...

518.4

...

0.3

...

Kapa HiFi HotStart DNA Pol

...

768

...

345.6

...

7.25

...

HPLC H2O

...

768

...

8352

...

11

...

Total Volume

...

768

...

12672

*Add 11 ul to each well of a hard shell, full skirt plate. Seal with bubble strips and store in refrigerator until needed label “NovaSeq4 PCR”. (If doing manually, one needs 2 ul of template and 2 ul of the primers).

...

Plate

...

16S MID

...

ITS MID

...

5CM1

...

16S0A1

...

N/A

...

16S0B1

...

N/A

...

5CM2

...

16S0C1

...

N/A

...

16S0D1

...

N/A

...

5CM3

...

16S0E1

...

N/A

...

16S0F1

...

N/A

...

5CM4

...

16S0G1

...

N/A

...

16S0H1

...

N/A

Run on Thermocycler Program GSAF36:

...

Temp C

...

Cycles

...

Time

...

95*

...

1X

...

3:00*

...

98

...

36X

...

0:30

...

62

...

36X

...

0:30

...

72

...

36X

...

0:30

...

72

...

1X

...

5:00

...

4

...

1X

...

0:00

MagBead Cleanup:

Some plates were cleaned up using the Nimbus platform protocol “AxyPrep MagBead PCR1 No MM”

Manually, it was done:

  • 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 (Plate name_PCR_MIDs)

qPCR NovaSeq5_Plate1

  • Make 1:1000 dilutions of column 3 from the PCR plates by adding 1 ul to 999 ul TE in a deep well plate:

...

 

...

1

...

2

...

3

...

4

...

5

...

6

...

7

...

8

...

9

...

10

...

11

...

12

...

A

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

NTC

...

NTC

...

NTC

...

B

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

0.0002 pM Std

...

0.0002 pM Std

...

0.0002 pM Std

...

C

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

0.002 pM Std

...

0.002 pM Std

...

0.002 pM Std

...

D

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

0.02 pM Std

...

0.02 pM Std

...

0.02 pM Std

...

E

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

0.2 pM Std

...

0.2 pM Std

...

0.2 pM Std

...

F

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

2 pM Std

...

2 pM Std

...

2 pM Std

...

G

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

20 pM Std

...

20 pM Std

...

20 pM Std

...

H

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

 

...

 

...

 

...

 

  • Make a 1:1000 dilution of HMax_PP Final Pool to check quality of pippin elution before sending out for RFS.

  • Add 16 ul of Illumina Library Quantification MasterMix to each well:

...

ul/rxn

...

Reagent

...

# of rxns

...

ul needed

...

10 ul

...

KAPA SYBR FAST qPCR MM (2X)

...

60

...

600

...

2 ul

...

Primer Premix (10X)

...

60

...

120

...

4 ul

...

Ultra Pure Water

...

60

...

240

...

16 ul

...

Total Volume

...

60

...

960

  • Add 4 ul of template, pool, or standards to each well:

...

 

...

1

...

2

...

3

...

4

...

5

...

6

...

7

...

8

...

9

...

10

...

11

...

12

...

A

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

HMax_PP_Final

...

 

...

NTC

...

NTC

...

NTC

...

B

...

5CM1_16S_Col3

...

5CM2_16S_Col3

...

5CM3_16S_Col3

...

5CM4_16S_Col3

...

5RM1_1nm_Pool

...

 

...

0.0002 pM Std

...

0.0002 pM Std

...

0.0002 pM Std

Info

Data was returned on 9-21-2021. Data was transferred to client and invoiced. M01247

5CM1 (16S only)

5CM2 (16S only)

5CM3 (16S only)

5CM4 (16S only)

MasterMix (make for 8 plates in 15mL conical tube)

ul/rxn

Reagent

# of rxns

ul needed (1.5X)

3

5X Kapa HiFi Buffer

768

3456

0.45

10M dNTPs

768

518.4

0.3

Kapa HiFi HotStart DNA Pol

768

345.6

7.25

HPLC H2O

768

8352

11

Total Volume

768

12672

*Add 11 ul to each well of a hard shell, full skirt plate. Seal with bubble strips and store in refrigerator until needed label “NovaSeq4 PCR”. (If doing manually, one needs 2 ul of template and 2 ul of the primers).

Plate

16S MID

ITS MID

5CM1

16S0A1

N/A

16S0B1

N/A

5CM2

16S0C1

N/A

16S0D1

N/A

5CM3

16S0E1

N/A

16S0F1

N/A

5CM4

16S0G1

N/A

16S0H1

N/A

Run on Thermocycler Program GSAF36:

Temp C

Cycles

Time

95*

1X

3:00*

98

36X

0:30

62

36X

0:30

72

36X

0:30

72

1X

5:00

4

1X

0:00

MagBead Cleanup:

Some plates were cleaned up using the Nimbus platform protocol “AxyPrep MagBead PCR1 No MM”

Manually, it was done:

  • 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 (Plate name_PCR_MIDs)

qPCR NovaSeq5_Plate1

  • Make 1:1000 dilutions of column 3 from the PCR plates by adding 1 ul to 999 ul TE in a deep well plate:

 

1

2

3

4

5

6

7

8

9

10

11

12

A

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

NTC

NTC

NTC

B

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

0.0002 pM Std

0.0002 pM Std

0.0002 pM Std

C

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

0.002 pM Std

0.002 pM Std

0.002 pM Std

D

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

0.02 pM Std

0.02 pM Std

0.02 pM Std

E

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

0.2 pM Std

0.2 pM Std

0.2 pM Std

F

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

2 pM Std

2 pM Std

2 pM Std

G

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

20 pM Std

20 pM Std

20 pM Std

H

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

 

 

 

  • Make a 1:1000 dilution of HMax_PP Final Pool to check quality of pippin elution before sending out for RFS.

  • Add 16 ul of Illumina Library Quantification MasterMix to each well:

ul/rxn

Reagent

# of rxns

ul needed

10 ul

KAPA SYBR FAST qPCR MM (2X)

60

600

2 ul

Primer Premix (10X)

60

120

4 ul

Ultra Pure Water

60

240

16 ul

Total Volume

60

960

  • Add 4 ul of template, pool, or standards to each well:

 

1

2

3

4

5

6

7

8

9

10

11

12

A

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

HMax_PP_Final

 

NTC

NTC

NTC

B

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

5RM1_1nm_Pool

 

0.0002 pM Std

0.0002 pM Std

0.0002 pM Std

C

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

0.002 pM Std

0.002 pM Std

0.002 pM Std

D

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

0.02 pM Std

0.02 pM Std

0.02 pM Std

E

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

0.2 pM Std

0.2 pM Std

0.2 pM Std

F

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

2 pM Std

2 pM Std

2 pM Std

G

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

20 pM Std

20 pM Std

20 pM Std

H

5CM1_16S_Col3

5CM2_16S_Col3

5CM3_16S_Col3

5CM4_16S_Col3

 

 

 

 

Results:

Average results for the following plates (column 3):

5CM1_16S: 6.03 nanomoles

5CM2_16S: 9.32 nanomoles

5CM3_16S: 3.52 nanomoles

5CM4_16S: 9.77 nanomoles

Results for HMax_PP_Final can be viewed in result report below and have been added to the HMax page here.

Full result report can be viewed below:

View file
name5CM_HMAXPP_5RM1.csv

Pooling and Shipping:

The client, Chris MacGlover, is concerned about the nasal swabs not sequencing as occurred with his last attempt. We will send 3 nasal, one excontrol, and one other sample to be fragment analyzed at CU:

5CM1 A4 (18-143N), 5CM1 H12 (SB33N), 5CM2 A4 ( 18-143T), 5CM3 H12 (51N), and 5CM1 A3 (excontrol)

Fragment Analysis returned from the Genomics Core lead us to allow sequencing to proceed.

...

Well

Conc pg/ul

Sample Description

EL1

2350

Electronic Ladder

A1

2380

18-143T

B1

1090

SB33N

C1

1390

18-143N

D1

1450

51N

E1

688

excontrol

F1

1890

5CM pool