Page Comparison

Prepare two ITS and two 16S PCR plates for the following template plates:

ALF_PLT1_2020_END	ALF_PLT2_2020_END	ALF_PLT3_2020_END	ALF_PLT4_2020_END
ALF_PLT5_2020_END	ALF_PLT6_2020_END	ALF_PLT7_2020_END	ALF_PLT8_2020_END
ALF_PLT1_2020_EPI	ALF_PLT2_2020_EPI	ALF_PLT3_2020_EPI	ALF_PLT4_2020_EPI
ALF_PLT5_2020_EPI	ALF_PLT6_2020_EPI	ALF_PLT7_2020_EPI	ALF_PLT8_2020_EPI
ALF_PLT9_2020_EPI	ALF_PLT1_2020_SOIL
ALF_PLT1_2017_GBS	ALF_PLT2_2017_GBS	ALF_PLT3_2017_GBS	ALF_PLT4_2017_GBS
ALF_PLT5_2017_GBS	ALF_PLT6_2017_GBS	ALF_PLT7_2017_GBS	ALF_PLT8_2017_GBS
ALF_PLT9_2017_GBS	ALF_PLT10_2017_GBS	ALF_PLT11_2017_GBS

MasterMix (make for 16 plates in 50mL conical tube)

ul/rxn	Reagent	# of rxns	ul needed
3	5X Kapa HiFi Buffer	1700	5100
0.45	10M dNTPs	1700	765
0.3	Kapa HiFi HotStart DNA Pol	1700	510
7.25	Ultra Pure H2O	1700	12,325
11	Total Volume	1700	18,700

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

Plates	16S MIDs	ITS MIDs
ALF_PLT1_2020_END	long16S0A1	longITS0A1
ALF_PLT1_2020_END	long16S0B1	longITS0B1
ALF_PLT2_2020_END	long16S0C1	longITS0C1
ALF_PLT2_2020_END	long16S0D1	longITS0D1
ALF_PLT3_2020_END	long16S0E1	longITS0E1
ALF_PLT3_2020_END	long16S0F1	longITS0F1
ALF_PLT4_2020_END	long16S0G1	longITS0G1
ALF_PLT4_2020_END	long16S0H1	longITS0H1

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”

...

MasterMix (make for 16 plates in 50mL conical tube)

ul/rxn	Reagent	# of rxns	ul needed
3	5X Kapa HiFi Buffer	1700	5100
0.45	10M dNTPs	1700	765
0.3	Kapa HiFi HotStart DNA Pol	1700	510
7.25	Ultra Pure H2O	1700	12,325
11	Total Volume	1700	18,700

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

Plates	16S MIDs	ITS MIDs
ALF_PLT5_2020_END	long16S0A2	longITS0A2
ALF_PLT5_2020_END	long16S0B2	longITS0B2
ALF_PLT6_2020_END	long16S0C2	longITS0C2
ALF_PLT6_2020_END	long16S0D2	longITS0D2
ALF_PLT7_2020_END	long16S0E2	longITS0E2
ALF_PLT7_2020_END	long16S0F2	longITS0F2
ALF_PLT8_2020_END	long16S0G2	longITS0G2
ALF_PLT8_2020_END	long16S0H2	longITS0H2

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”

...

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

	1	2	3	4	11	12
A	ALF_END_PLT1_16S_3B	ALF_END_PLT3_16S_3B	ALF_END_PLT5_16S_3B	ALF_END_PLT7_16S_3B		NTC
B	ALF_END_PLT1_16S_9H	ALF_END_PLT3_16S_9H	ALF_END_PLT5_16S_9H	ALF_END_PLT7_16S_9H	04.0002 pM Std	0.0002 pM Std
C	ALF_END_PLT1_ITS_3B	ALF_END_PLT5_ITS_3B	ALF_END_PLT3_ITS_3B	ALF_END_PLT7_ITS_3B	0.002 pM Std	0.002 pM Std
D	ALF_END_PLT1_ITS_9H	ALF_END_PLT5_ITS_9H	ALF_END_PLT3_ITS_9H	ALF_END_PLT7_ITS_9H	0.02 pM Std	0.02 pM Std
E	ALF_END_PLT2_16S_3B	ALF_END_PLT4_16S_3B	ALF_END_PLT6_16S_3B	ALF_END_PLT8_16S_3B	0.2 pM Std	0.2 pM Std
F	ALF_END_PLT2_16S_9H	ALF_END_PLT4_16S_9H	ALF_END_PLT6_16S_9H	ALF_END_PLT8_16S_9H	2 pM Std	2 pM Std
G	ALF_END_PLT2_ITS_3B	ALF_END_PLT4_ITS_3B	ALF_END_PLT6_ITS_3B	ALF_END_PLT8_ITS_3B	20 pM Std	20 pM Std
H	ALF_END_PLT2_ITS_9H	ALF_END_PLT4_ITS_9H	ALF_END_PLT6_ITS_9H	ALF_END_PLT8_ITS_9H

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	10	11	12
A	ALF_END_PLT1_16S_3B	ALF_END_PLT3_16S_3B	ALF_END_PLT5_16S_3B	ALF_END_PLT7_16S_3B	NTC	NTC	NTC
B	ALF_END_PLT1_16S_9H	ALF_END_PLT3_16S_9H	ALF_END_PLT5_16S_9H	ALF_END_PLT7_16S_9H	0.0002 pM Std	0.0002 pM Std	0.0002 pM Std
C	ALF_END_PLT1_ITS_3B	ALF_END_PLT5_ITS_3B	ALF_END_PLT3_ITS_3B	ALF_END_PLT7_ITS_3B	0.002 pM Std	0.002 pM Std	0.002 pM Std
D	ALF_END_PLT1_ITS_9H	ALF_END_PLT5_ITS_9H	ALF_END_PLT3_ITS_9H	ALF_END_PLT7_ITS_9H	0.02 pM Std	0.02 pM Std	0.02 pM Std
E	ALF_END_PLT2_16S_3B	ALF_END_PLT4_16S_3B	ALF_END_PLT6_16S_3B	ALF_END_PLT8_16S_3B	0.2 pM Std	0.2 pM Std	0.2 pM Std
F	ALF_END_PLT2_16S_9H	ALF_END_PLT4_16S_9H	ALF_END_PLT6_16S_9H	ALF_END_PLT8_16S_9H	2 pM Std	2 pM Std	2 pM Std
G	ALF_END_PLT2_ITS_3B	ALF_END_PLT4_ITS_3B	ALF_END_PLT6_ITS_3B	ALF_END_PLT8_ITS_3B	20 pM Std	20 pM Std	20 pM Std
H	ALF_END_PLT2_ITS_9H	ALF_END_PLT4_ITS_9H	ALF_END_PLT6_ITS_9H	ALF_END_PLT8_ITS_9H

Results:

Average results for the following plates:

...

MasterMix (make for 16 plates in 50mL conical tube)

ul/rxn	Reagent	# of rxns	ul needed
3	5X Kapa HiFi Buffer	1700	5100
0.45	10M dNTPs	1700	765
0.3	Kapa HiFi HotStart DNA Pol	1700	510
7.25	Ultra Pure H2O	1700	12,325
11	Total Volume	1700	18,700

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

Plates	16S MIDs	ITS MIDs
ALF_PLT1_2020_EPI	long16S0A3	longITS0A3
ALF_PLT1_2020_EPI	long16S0B3	longITS0B3
ALF_PLT2_2020_EPI	long16S0C3	longITS0C3
ALF_PLT2_2020_EPI	long16S0D3	longITS0D3
ALF_PLT3_2020_EPI	long16S0E3	longITS0E3
ALF_PLT3_2020_EPI	long16S0F3	longITS0F3
ALF_PLT4_2020_EPI	long16S0G3	longITS0G3
ALF_PLT4_2020_EPI	long16S0H3	longITS0H3

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”

...

MasterMix (make for 16 plates in 50mL conical tube)

ul/rxn	Reagent	# of rxns	ul needed
3	5X Kapa HiFi Buffer	1700	5100
0.45	10M dNTPs	1700	765
0.3	Kapa HiFi HotStart DNA Pol	1700	510
7.25	Ultra Pure H2O	1700	12,325
11	Total Volume	1700	18,700

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

Plates	16S MIDs	ITS MIDs
ALF_PLT5_2020_EPI	long16S0A4	longITS0A4
ALF_PLT5_2020_EPI	long16S0B4	longITS0B4
ALF_PLT6_2020_EPI	long16S0C4	longITS0C4
ALF_PLT6_2020_EPI	long16S0D4	longITS0D4
ALF_PLT7_2020_EPI	long16S0E4	longITS0E4
ALF_PLT7_2020_EPI	long16S0F4	longITS0F4
ALF_PLT8_2020_EPI	long16S0G4	longITS0G4
ALF_PLT8_2020_EPI	long16S0H4	longITS0H4

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”

...

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

	5	6	7	8	10	11	12
A	ALF_EPI_PLT1_16S_3B	ALF_EPI_PLT3_16S_3B	ALF_EPI_PLT5_16S_3B	ALF_EPI_PLT7_16S_3B	NTC	NTC	NTC
B	ALF_EPI_PLT1_16S_9H	ALF_EPI_PLT3_16S_9H	ALF_EPI_PLT5_16S_9H	ALF_EPI_PLT7_16S_9H	0.0002 pM Std	0.0002 pM Std	0.0002 pM Std
C	ALF_EPI_PLT1_ITS_3B	ALF_EPI_PLT3_ITS_3B	ALF_EPI_PLT5_ITS_3B	ALF_EPI_PLT7_ITS_3B	0.002 pM Std	0.002 pM Std	0.002 pM Std
D	ALF_EPI_PLT1_ITS_9H	ALF_EPI_PLT3_ITS_9H	ALF_EPI_PLT5_ITS_9H	ALF_EPI_PLT7_ITS_9H	0.02 pM Std	0.02 pM Std	0.02 pM Std
E	ALF_EPI_PLT2_16S_3B	ALF_EPI_PLT4_16S_3B	ALF_EPI_PLT6_16S_3B	ALF_EPI_PLT8_16S_3B	0.2 pM Std	0.2 pM Std	0.2 pM Std
F	ALF_EPI_PLT2_16S_9H	ALF_EPI_PLT4_16S_9H	ALF_EPI_PLT6_16S_9H	ALF_EPI_PLT8_16S_9H	2 pM Std	2 pM Std	2 pM Std
G	ALF_EPI_PLT2_ITS_3B	ALF_EPI_PLT4_ITS_3B	ALF_EPI_PLT6_ITS_3B	ALF_EPI_PLT8_ITS_3B	20 pM Std	20 pM Std	20 pM Std
H	ALF_EPI_PLT2_ITS_9H	ALF_EPI_PLT4_ITS_9H	ALF_EPI_PLT6_ITS_9H	ALF_EPI_PLT8_ITS_9H

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:

...

MasterMix (make for 16 plates in 50mL conical tube)

ul/rxn	Reagent	# of rxns	ul needed
3	5X Kapa HiFi Buffer	1700	5100
0.45	10M dNTPs	1700	765
0.3	Kapa HiFi HotStart DNA Pol	1700	510
7.25	Ultra Pure H2O	1700	12,325
11	Total Volume	1700	18,700

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

Plates	16S MIDs	ITS MIDs
ALF_PLT1_2017_GBS	long16S0A5	longITS0A5
ALF_PLT1_2017_GBS	long16S0B5	longITS0B5
ALF_PLT2_2017_GBS	long16S0C5	longITS0C5
ALF_PLT2_2017_GBS	long16S0D5	longITS0D5
ALF_PLT3_2017_GBS	long16S0E5	longITS0E5
ALF_PLT3_2017_GBS	long16S0F5	longITS0F5
ALF_PLT4_2017_GBS	long16S0G5	longITS0G5
ALF_PLT4_2017_GBS	long16S0H5	longITS0H5

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”

...

MasterMix (make for 12 plates in 50mL conical tube)

ul/rxn	Reagent	# of rxns	ul needed
3	5X Kapa HiFi Buffer	1700	5100
0.45	10M dNTPs	1700	765
0.3	Kapa HiFi HotStart DNA Pol	1700	510
7.25	Ultra Pure H2O	1700	12,325
11	Total Volume	1700	18,700

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

Plates	16S MIDs	ITS MIDs
ALF_PLT5_2017_GBS	long16S0A6	longITS0A6
ALF_PLT5_2017_GBS	long16S0B6	longITS0B6
ALF_PLT6_2017_GBS	long16S0C6	longITS0C6
ALF_PLT6_2017_GBS	long16S0D6	longITS0D6
ALF_PLT7_2017_GBS	long16S0E6	longITS0E6
ALF_PLT7_2017_GBS	long16S0F6	longITS0F6
ALF_PLT8_2017_GBS	long16S0G6	longITS0G6
ALF_PLT8_2017_GBS	long16S0H6	longITS0H6

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”

...

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)

MasterMix (

...

in 50mL conical tube)

ul/rxn	Reagent	# of rxns	ul needed
3	5X Kapa HiFi Buffer

2100

1550

6,300

4650
0.45	10M dNTPs

2100

1550

945

697.5
0.3	Kapa HiFi HotStart DNA Pol

2100

1550

630

465
7.25	Ultra Pure H2O

2100

1550

15

11,

225

237.5
11	Total Volume

2100

1550

23

17,

100

050

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

Plates	16S MIDs	ITS MIDs
ALF_PLT9_2017_GBS	long16S0A7	longITS0A7
ALF_PLT9_2017_GBS	long16S0B7	longITS0B7
ALF_PLT10_2017_GBS	long16S0C7	longITS0C7
ALF_PLT10_2017_GBS	long16S0D7	longITS0D7
ALF_PLT11_2017_GBS	long16S0E7	longITS0E7
ALF_PLT11_2017_GBS	long16S0F7	longITS0F7
ALF_PLT9_2020_EPI	long16S0G7	longITS0G7
ALF_PLT9_2020_EPI	long16S0H7	longITS0H7
ALF_PLT1_2020_SOIL	long16S0A8	longITS0A8
ALF_PLT1_2020_SOIL	long16S0B8	longITS0B8

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”

...

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

	1	2	3	4	5	6	7	10	11	12
A	ALF_GBS_PLT1_16S_3B	ALF_GBS_PLT3_16S_3B	ALF_GBS_PLT5_16S_3B	ALF_GBS_PLT7_16S_3B	ALF_GBS_PLT9_16S_3B	ALF_GBS_PLT11_16S_3B	ALF_PLT1_2020_SOIL_16S_3B	NTC	NTC	NTC
B	ALF_GBS_PLT1_16S_9H	ALF_GBS_PLT3_16S_9H	ALF_GBS_PLT5_16S_9H	ALF_GBS_PLT7_16S_9H	ALF_GBS_PLT9_16S_9H	ALF_GBS_PLT11_16S_9A	ALF_PLT1_2020_SOIL_16S_9H	0.0002 pM Std	0.0002 pM Std	0.0002 pM Std
C	ALF_GBS_PLT1_ITS_3B	ALF_GBS_PLT3_ITS_3B	ALF_GBS_PLT5_ITS_3B	ALF_GBS_PLT7_ITS_3B	ALF_GBS_PLT9_ITS_3B	ALF_GBS_PLT11_ITS_3B	ALF_PLT1_2020_SOIL_ITS_3B	0.002 pM Std	0.002 pM Std	0.002 pM Std
D	ALF_GBS_PLT1_ITS_9H	ALF_GBS_PLT3_ITS_9H	ALF_GBS_PLT5_ITS_9H	ALF_GBS_PLT7_ITS_9H	ALF_GBS_PLT9_ITS_9H	ALF_GBS_PLT11_ITS_9A	ALF_PLT1_2020_SOIL_ITS_9H	0.02 pM Std	0.02 pM Std	0.02 pM Std
E	ALF_GBS_PLT2_16S_3B	ALF_GBS_PLT4_16S_3B	ALF_GBS_PLT6_16S_3B	ALF_GBS_PLT8_16S_3B	ALF_GBS_PLT10_16S_3B	ALF_PLT9_2020_EPI_16S_1B		0.2 pM Std	0.2 pM Std	0.2 pM Std
F	ALF_GBS_PLT2_16S_9H	ALF_GBS_PLT4_16S_9H	ALF_GBS_PLT6_16S_9H	ALF_GBS_PLT8_16S_9H	ALF_GBS_PLT10_16S_9H	ALF_PLT9_2020_EPI_16S_3H		2 pM Std	2 pM Std	2 pM Std
G	ALF_GBS_PLT2_ITS_3B	ALF_GBS_PLT4_ITS_3B	ALF_GBS_PLT6_ITS_3B	ALF_GBS_PLT8_ITS_3B	ALF_GBS_PLT10_ITS_3B	ALF_PLT9_2020_EPI_ITS_1B		20 pM Std	20 pM Std	20 pM Std
H	ALF_GBS_PLT2_ITS_9H	ALF_GBS_PLT4_ITS_9H	ALF_GBS_PLT6_ITS_9H	ALF_GBS_PLT8_ITS_9H	ALF_GBS_PLT10_ITS_9H	ALF_PLT9_2020_EPI_ITS_3H

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

90	600
2 ul	Primer Premix (10X)

60

90	120
4 ul	Ultra Pure Water

60

90	240
16 ul	Total Volume

60

90

960

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

	1	2	3	4	5	6	7	8	10	11	12
A	ALF_GBS_PLT1_16S_3B	ALF_GBS_PLT3_16S_3B	ALF_GBS_PLT5_16S_3B	ALF_GBS_PLT7_16S_3B	ALF_GBS_PLT9_16S_3B	ALF_GBS_PLT11_16S_3B	ALF_PLT1_2020_SOIL_16S_3B	TRNL_16S_Pool	NTC	NTC	NTC
B	ALF_GBS_PLT1_16S_9H	ALF_GBS_PLT3_16S_9H	ALF_GBS_PLT5_16S_9H	ALF_GBS_PLT7_16S_9H	ALF_GBS_PLT9_16S_9H	ALF_GBS_PLT11_16S_9A	ALF_PLT1_2020_SOIL_16S_9H	TRNL_16S_Pool	0.0002 pM Std	0.0002 pM Std	0.0002 pM Std
C	ALF_GBS_PLT1_ITS_3B	ALF_GBS_PLT3_ITS_3B	ALF_GBS_PLT5_ITS_3B	ALF_GBS_PLT7_ITS_3B	ALF_GBS_PLT9_ITS_3B	ALF_GBS_PLT11_ITS_3B	ALF_PLT1_2020_SOIL_ITS_3B		0.002 pM Std	0.002 pM Std	0.002 pM Std
D	ALF_GBS_PLT1_ITS_9H	ALF_GBS_PLT3_ITS_9H	ALF_GBS_PLT5_ITS_9H	ALF_GBS_PLT7_ITS_9H	ALF_GBS_PLT9_ITS_9H	ALF_GBS_PLT11_ITS_9A	ALF_PLT1_2020_SOIL_ITS_9H		0.02 pM Std	0.02 pM Std	0.02 pM Std
E	ALF_GBS_PLT2_16S_3B	ALF_GBS_PLT4_16S_3B	ALF_GBS_PLT6_16S_3B	ALF_GBS_PLT8_16S_3B	ALF_GBS_PLT10_16S_3B	ALF_PLT9_2020_EPI_16S_1B	TRNL_T_Pool		0.2 pM Std	0.2 pM Std	0.2 pM Std
F	ALF_GBS_PLT2_16S_9H	ALF_GBS_PLT4_16S_9H	ALF_GBS_PLT6_16S_9H	ALF_GBS_PLT8_16S_9H	ALF_GBS_PLT10_16S_9H	ALF_PLT9_2020_EPI_16S_3H	TRNL_T_Pool		2 pM Std	2 pM Std	2 pM Std
G	ALF_GBS_PLT2_ITS_3B	ALF_GBS_PLT4_ITS_3B	ALF_GBS_PLT6_ITS_3B	ALF_GBS_PLT8_ITS_3B	ALF_GBS_PLT10_ITS_3B	ALF_PLT9_2020_EPI_ITS_1B	TRNL_T_Pool		20 pM Std	20 pM Std	20 pM Std
H	ALF_GBS_PLT2_ITS_9H	ALF_GBS_PLT4_ITS_9H	ALF_GBS_PLT6_ITS_9H	ALF_GBS_PLT8_ITS_9H	ALF_GBS_PLT10_ITS_9H	ALF_PLT9_2020_EPI_ITS_3H	TRNL_16S_Pool

Results:

Average results for the following plates:

ALF_GBS_PLT1_16S: 11.05 nanomoles

ALF_GBS_PLT1_ITS: 30.57 nanomoles

ALF_GBS_PLT2_16S: 14.58 nanomoles

ALF_GBS_PLT2_ITS: 6.37 nanomoles

ALF_GBS_PLT3_16S: 18.19 nanomoles

ALF_GBS_PLT3_ITS: 6.43 nanomoles

ALF_GBS_PLT4_16S: 6.71 nanomoles

ALF_GBS_PLT4_ITS: 5.98 nanomoles

ALF_GBS_PLT5_16S: 7.01 nanomoles

ALF_GBS_PLT5 ITS: 5.84 nanomoles

ALF_GBS_PLT5PLT6_16S: ALF_GBS_PLT5_ITS:: 6.29 nanomoles

ALF_GBS_PLT6_16S:ALF_GBS_PLT6_ITS: 1.22 nanomoles

ALF_GBS_PLT7_16S: 1.96 nanomoles

ALF_GBS_PLT7_ITS: 2.77 nanomoles

ALF_GBS_PLT8_16S: 9.72 nanomoles

ALF_GBS_PLT8_ITS: 6.96 nanomoles

ALF_GBS_PLT9_16S: 13.38 nanomoles

ALF_GBS_PLT9_ITS: 15.33 nanomoles

ALF_GBS_PLT10_16S: 8.09 nanomoles

ALF_GBS_PLT10_ITS: 8.1 nanomoles

ALF_GBS_PLT11_16S: 6.30 nanomoles

ALF_GBS_PLT11_ITS: 7.88 nanomoles

ALF_PLT9_2020_EPI_16S: 1.69 nanomoles

ALF_PLT9_2020_EPI_ITS: 0.677 nanomoles

ALF_PLT1_2020_SOIL_16S: 3.29 nanomoles

ALF_PLT1_2020_SOIL_ITS: 12.42 nanomoles

The full result report can be viewed below:

View file

name	ALF_TRNL_16S_ITS.csv

Versions Compared

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New Version Current

Key

MasterMix (make for 16 plates in 50mL conical tube)

MagBead Cleanup:

MasterMix (make for 16 plates in 50mL conical tube)

MagBead Cleanup:

Results:

MasterMix (make for 16 plates in 50mL conical tube)

MagBead Cleanup:

MasterMix (make for 16 plates in 50mL conical tube)

MagBead Cleanup:

MasterMix (make for 16 plates in 50mL conical tube)

MagBead Cleanup:

MasterMix (make for 12 plates in 50mL conical tube)

MagBead Cleanup:

MasterMix (

in 50mL conical tube)

MagBead Cleanup:

Results: