| Info |
|---|
| Info |
Plates in red have been extracted, PCRed, cleaned, quantified, and normalized by 7-15-22. Bolded red have been qPCRed. |
NS6 Plates
6AC1 | 6AC2 | 6AC3 | |
6BM1 | 6BM2 | ||
6LVD1 | 6LVD2 | 6LVD3 | 6LVD4 |
6NW1 | 6NW2 | 6NW3 | 6NW4 |
6RD1 | 6RD2 | 6RD3 | |
6GTL1 | 6GTL2 | ||
6RD4 | 6RD5 | 6RD6 | 6RD7 |
6LVD5 |
|
| |
MP002 | MP003 | MP006 |
Current total number of plates for ITS/16S PCR: 20 + 1 plate repeat (sm19-fire)
MasterMix (make for 16 plates in 50mL conical tube)
...
Add 11 ul to each well of a hard shell, full skirt plate. Seal with bubble strips and store in refrigerator until needed label “NS5 “NS6 PCR”. (If doing manually, one needs 2 ul of template and 2 ul of the primers).
...
Plate | 16S Primer | ITS Primer |
|---|---|---|
6LVD4 | 16S0A3 | ITS0A3 |
16S0B3 | ITS0B3 | |
6RD1 | 16S0C3 | ITS0C3 |
16S0D3 | ITS0D3 | |
6RD2 | 16S0E3 | ITS0E3 |
16S0F3 | ITS0F3 | |
6NW16RD3 | 16S0G3 | ITS0G3 |
16S0H3 | ITS0H3 |
Run on Thermocycler Program GSAF36:
...
Add 11 ul to each well of a hard shell, full skirt plate. Seal with bubble strips and store in refrigerator until needed labeled “NS5 PCR”. (If doing manually, one needs 2 ul of template and 2 ul of the primers).
...
Plate
...
16S Primer
...
ITS Primer
...
6NW2
...
16S0A4
...
ITS0A4
...
16S0B4
...
ITS0B4
...
6NW3
...
16S0C4
...
ITS0C4
...
16S0D4
...
ITS0D4
...
6NW4
...
16S0E4
...
ITS0E4
...
16S0F4
...
Plate | 16S Primer | ITS Primer |
|---|---|---|
6GTL1 | 16S0G4 | ITS0G4 |
16S0H4 | ITS0H4 | |
6GTL2 | 16S0A5 | ITS0A5 |
16S0B5 | ITS0B5 |
...
Add 16 ul of Illumina Library Quantification MasterMix to each well:
1
2
3
4
5
6
7
8
9
10
11
12
A
6AC3_ITS
6AC3_16S
6RD1_ITS
6RD1_16S
6BM1_ITS
6BM1_16S
6LVD1_ITS
6LVD1_16S
6GTL1_ITS
6GTL1_16S
20 pM Std
20 pM Std
B
6AC3_ITS
6AC3_16S
6RD1_ITS
6RD1_16S
6BM1_ITS
6BM1_16S
6LVD1_ITS
6LVD1_16S
6GTL1_ITS
6GTL1_16S
2 pM Std
2 pM Std
ul/rxn | Reagent | # of rxns | ul needed |
|---|---|---|---|
10 ul | KAPA SYBR FAST qPCR MM (2X) | 110 | 1100 |
2 ul | Primer Premix (10X) | 110 | 220 |
4 ul | Ultra Pure Water | 110 | 440 |
16 ul | Total Volume | 110 | 1760 |
Add 4 ul of template, pool, or standards to each well:
110 | 220 | ||
4 ul | Ultra Pure Water | 110 | 440 |
16 ul | Total Volume | 110 | 1760 |
Add 4 ul of template, pool, or standards to each well:
Plate Name
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | ||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
A | 6AC3_ITS | 6AC3_16S | 6RD1_ITS | 6RD1_16S | 6BM1_ITS | 6BM1_16S | 6LVD1_ITS | 6LVD1_16S | 6GTL1_ITS | 6GTL1_16S | 0.2 20 pM Std0.2 | 20 pM Std | ||
DB | 6AC3_ITS | 6AC3_16S | 6RD1_ITS | 6RD1_16S | 6BM1_ITS | 6BM1_16S | 6LVD1_ITS | 6LVD1_16S | 6GTL1_ITS | 6GTL1_16S | 0.02 2 pM Std0.02 | 2 pM Std | ||
EC | 6AC3_ITS | 6AC3_16S | 6RD1_ITS | 6RD1_16S | 6BM1_ITS | 6BM1_16S | 6LVD1_ITS | 6LVD1_16S | 6GTL1_ITS | 6GTL1_16S | 0.002 2 pM Std | 0.002 2 pM Std | ||
FD | 6AC3_ITS | 6AC3_16S | 6RD1_ITS | 6RD1_16S | 6BM1_ITS | 6BM1_16S | 6LVD1_ITS | 6LVD1_16S | 6GTL1_ITS | 6GTL1_16S | 0.0002 02 pM Std | 0.0002 02 pM Std | ||
GE | 6AC3_ITS | 6AC3_16S | 6RD1_ITS | 6RD1_16S | 6BM1_ITS | 6BM1_16S | 6LVD1_ITS | 6LVD1_16S | 6GTL1_ITS | 6GTL1_16S | NTC | NTC | H0.002 pM Std | 0.002 pM Std |
F | 6AC3_ITS | 6AC3_16S | 6RD1_ITS | 6RD1_16S | 6BM1_ITS | 6BM1_16S | 6LVD1_ITS | 6LVD1_16S | 6GTL1_ITS | 6GTL1_16S | NTC | NTC |
Results:
0.0002 pM Std | 0.0002 pM Std |
G | 6AC3_ITS |
4.125
6AC3_16S |
6RD1_ITS |
10.152
6RD1_16S |
15.709
6BM1_ITS |
7.874
6BM1_16S |
6LVD1_ITS |
4.532
6LVD1_16S |
6GTL1_ITS |
5.992
6GTL1_16S |
Needs to be redone
Full qPCR results below:
| View file | ||
|---|---|---|
|
qPCR NovaSeq6_Set3
Make 1:1000 dilutions of column 8 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
6LVD2_ITS
6LVD2_16S
2TRNL3_16S
2TRNL3_TRNL
2TRNL2_TRNL
2TRNL2_16S
6RD2_16S
6RD2_ITS
6GTL1_16S
2TRNL1_16S_2
NTC
B
6LVD2_ITS
6LVD2_16S
2TRNL3_16S
2TRNL3_TRNL
2TRNL2_TRNL
2TRNL2_16S
6RD2_16S
6RD2_ITS
6GTL1_16S
2TRNL1_16S_2
04.0002 pM Std
0.0002 pM Std
C
6LVD2_ITS
6LVD2_16S
2TRNL3_16S
2TRNL3_TRNL
2TRNL2_TRNL
2TRNL2_16S
6RD2_16S
6RD2_ITS
6GTL1_16S
2TRNL1_16S_2
0.002 pM Std
0.002 pM Std
NTC | NTC | |||||||||||
H | 6AC3_ITS | 6AC3_16S | 6RD1_ITS | 6RD1_16S | 6BM1_ITS | 6BM1_16S | 6LVD1_ITS | 6LVD1_16S | 6GTL1_ITS | 6GTL1_16S | NTC | NTC |
Results:
Plate Name | Average Result (nanomoles) |
|---|---|
6AC3_ITS | 4.125 |
6AC3_16S | 7.46 |
6RD1_ITS | 10.152 |
6RD1_16S | 15.709 |
6BM1_ITS | 7.874 |
6BM1_16S | 4.842 |
6LVD1_ITS | 4.532 |
6LVD1_16S | 8.102 |
6GTL1_ITS | 5.992 |
6GTL1_16S | Needs to be redone |
Full qPCR results below:
| View file | ||
|---|---|---|
|
qPCR NovaSeq6_Set3
Make 1:1000 dilutions of column 8 from the PCR plates by adding 1 ul to 999 ul TE in a deep well plate:
Add 4 ul of template, pool, or standards to each well:
1
2
3
4
5
6
7
8
9
10
11
12
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | ||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
A | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | 0.02 pM Std | 0.02 pM Std | E | NTC |
B | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | 004.2 0002 pM Std | 0.2 0002 pM Std | ||
FC | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | 2 0.002 pM Std | 2 0.002 pM Std | ||
GD | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | 20 0.02 pM Std | 20 0.02 pM Std | ||
HE | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S2TRNL1_16S_2 |
|
|
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)
...
110
...
1100
...
2 ul
...
Primer Premix (10X)
...
110
...
220
...
4 ul
...
Ultra Pure Water
...
110
...
440
...
16 ul
...
Total Volume
...
110
...
1760
2TRNL1_16S_2 | 0.2 pM Std | 0.2 pM Std | ||||||||||
F | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | 2 pM Std | 2 pM Std |
G | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | 20 pM Std | 20 pM Std |
H | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S |
2 pM Std
2 pM Std
C
6LVD2_ITS
6LVD2_16S
2TRNL3_16S
2TRNL3_TRNL
2TRNL2_TRNL
2TRNL2_16S
6RD2_16S
6RD2_ITS
6GTL1_16S
2TRNL1_16S_2
0.2 pM Std
0.2 pM Std
_2 |
|
|
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) | 110 | 1100 |
2 ul | Primer Premix (10X) | 110 | 220 |
4 ul | Ultra Pure Water | 110 | 440 |
16 ul | Total Volume | 110 | 1760 |
Add 4 ul of template, pool, or standards to each well:
Plate Name
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | ||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
A | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | 0.02 20 pM Std0.02 | 20 pM Std | ||
EB | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | 0.002 2 pM Std0.002 | 2 pM Std | ||
FC | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | 0.0002 2 pM Std | 0.0002 2 pM Std | ||
GD | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | NTC | NTC | H0.02 pM Std | 0.02 pM Std |
E | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | NTC | NTC |
Results:
0.002 pM Std | 0.002 pM Std |
F | 6LVD2_ITS |
16.09
6LVD2_16S |
2TRNL3_16S |
2TRNL3_TRNL |
2TRNL2_TRNL |
2TRNL2_16S |
6RD2_16S |
28.48
6RD2_ITS |
29.45
6GTL1_16S |
2TRNL1_16S_2 |
0. |
Full qPCR results below:
...
0002 pM Std | 0.0002 pM Std | ||||
G | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_ |
...
qPCR NovaSeq6_Set4
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
...
...
...
NTC
...
B
...
...
04.0002 pM Std
...
0.0002 pM Std
...
C
...
...
0.002 pM Std
...
0.002 pM Std
...
D
...
...
0.02 pM Std
...
0.02 pM Std
...
E
...
...
0.2 pM Std
...
0.2 pM Std
...
F
...
...
2 pM Std
...
2 pM Std
...
G
...
...
20 pM Std
...
20 pM Std
...
H
...
...
...
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)
...
110
...
1100
...
2 ul
...
Primer Premix (10X)
...
110
...
220
...
4 ul
...
Ultra Pure Water
...
110
...
440
...
16 ul
...
Total Volume
...
110
...
1760
Add 4 ul of template, pool, or standards to each well:
1
2
3
4
5
6
7
8
9
10
11
12
A
NTC
NTC
NTC
B
TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | NTC | NTC | |||||
H | 6LVD2_ITS | 6LVD2_16S | 2TRNL3_16S | 2TRNL3_TRNL | 2TRNL2_TRNL | 2TRNL2_16S | 6RD2_16S | 6RD2_ITS | 6GTL1_16S | 2TRNL1_16S_2 | NTC | NTC |
Results:
Plate Name | Average Result (nanomoles) |
|---|---|
6LVD2_ITS | 16.09 |
6LVD2_16S | 21.71 |
2TRNL3_16S | 42.69 |
2TRNL3_TRNL | 8.68 |
2TRNL2_TRNL | 7.51 |
2TRNL2_16S | 36.27 |
6RD2_16S | 28.48 |
6RD2_ITS | 29.45 |
6GTL1_16S | 8.2 |
2TRNL1_16S_2 | 32.72 |
Full qPCR results below:
| View file | ||
|---|---|---|
|
qPCR NovaSeq6_Set4
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 | 6RD4_16S_Col3 | 6RD4_ITS_Col3 | 6RD5_16S_Col3 | 6RD5_ITS_Col3 | 6RD6_16S_Col3 | 6RD6_ITS_Col3 | 6RD7_16S_Col3 | 6RD7_ITS_Col3 | 6LVD5_16S_A1 |
|
| NTC |
B | 6RD4_16S_Col3 | 6RD4_ITS_Col3 | 6RD5_16S_Col3 | 6RD5_ITS_Col3 | 6RD6_16S_Col3 | 6RD6_ITS_Col3 | 6RD7_16S_Col3 | 6RD7_ITS_Col3 | 6LVD5_16S_B1 |
| 04.0002 pM Std | 0.0002 pM Std |
0.0002 pM Std
C
C | 6RD4_16S_Col3 | 6RD4_ITS_Col3 | 6RD5_16S_Col3 | 6RD5_ITS_Col3 | 6RD6_16S_Col3 | 6RD6_ITS_Col3 | 6RD7_16S_Col3 | 6RD7_ITS_Col3 | 6LVD5_16S_C1 |
| 0.002 pM Std | 0.002 pM Std |
D |
0.02 pM Std
0.02 pM Std
0.02 pM Std
E
0.2 pM Std
6RD4_16S_Col3 | 6RD4_ITS_Col3 | 6RD5_16S_Col3 | 6RD5_ITS_Col3 | 6RD6_16S_Col3 | 6RD6_ITS_Col3 | 6RD7_16S_Col3 | 6RD7_ITS_Col3 | 6LVD5_16S_D1 |
| 0.02 pM Std | 0. |
Full qPCR results below:
qPCR NovaSeq6_Set5
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
NTC
B
04.0002 pM Std
0.0002 pM Std
C
0.002 pM Std
0.002 pM Std
D
0.02 pM Std
0.02 pM Std
E
0.2 pM Std
0.2 pM Std
F
2 pM Std
2 pM Std
G
20 pM Std
20 pM Std
H
02 pM Std |
F
2 pM Std
2 pM Std
2 pM Std
G
20 pM Std
20 pM Std
20 pM Std
H
Results:
...
Plate Name
...
Average Result (nanomoles)
E | 6RD4_16S_Col3 | 6RD4_ITS_Col3 | 6RD5_16S_Col3 | 6RD5_ITS_Col3 | 6RD6_16S_Col3 | 6RD6_ITS_Col3 | 6RD7_16S_Col3 | 6RD7_ITS_Col3 | 6LVD5_ITS_A8 |
| 0.2 pM Std | 0.2 pM Std |
F | 6RD4_16S_Col3 | 6RD4_ITS_Col3 | 6RD5_16S_Col3 | 6RD5_ITS_Col3 | 6RD6_16S_Col3 | 6RD6_ITS_Col3 | 6RD7_16S_Col3 | 6RD7_ITS_Col3 | 6LVD5_ITS_B8 |
| 2 pM Std | 2 pM Std |
G | 6RD4_16S_Col3 | 6RD4_ITS_Col3 | 6RD5_16S_Col3 | 6RD5_ITS_Col3 | 6RD6_16S_Col3 | 6RD6_ITS_Col3 | 6RD7_16S_Col3 | 6RD7_ITS_Col3 | 6LVD5_ITS_C8 |
| 20 pM Std | 20 pM Std |
H | 6RD4_16S_Col3 | 6RD4_ITS_Col3 | 6RD5_16S_Col3 | 6RD5_ITS_Col3 | 6RD6_16S_Col3 | 6RD6_ITS_Col3 | 6RD7_16S_Col3 | 6RD7_ITS_Col3 | 6LVD5_ITS_D8 |
|
|
|
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) | 110 | 1100 |
2 ul | Primer Premix (10X) | 110 | 220 |
4 ul | Ultra Pure Water | 110 | 440 |
16 ul | Total Volume | 110 | 1760 |
Add 4 ul of template, pool, or standards to each well:
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
|---|
11
12
A
NTC
NTC
NTC
B
0.0002 pM Std
0.0002 pM Std
0.0002 pM Std
C
0.002 pM Std
0.002 pM Std
0.002 pM Std
D
0.02 pM Std
0.02 pM Std
0.02 pM Std
E
0.2 pM Std
0.2 pM Std
0.2 pM Std
F
2 pM Std
2 pM Std
2 pM Std
G
20 pM Std
20 pM Std
20 pM Std
H
Results:
...
Plate Name
...
Average Result (nanomoles)
Full qPCR results below:
qPCR NovaSeq6_Set6
Make 1:1000 dilutions of column 3 from the PCR plates by adding 1 ul to 999 ul TE in a deep well plate:
...
G
...
20 pM Std
...
20 pM Std
...
20 pM Std
...
H
...
...
...
Results:
...
Plate Name
...
Average Result (nanomoles)
...
1
2
3
4
5
6
7
8
9
10
11
12
A
NTC
B
04.0002 pM Std
0.0002 pM Std
C
0.002 pM Std
0.002 pM Std
D
11 | 12 | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
A | 6RD4_16S | 6RD4_ITS | 6RD5_16S | 6RD5_ITS | 6RD6_16S | 6RD6_ITS | 6RD7_16S | 6RD7_ITS | 6LVD5_16S | NTC | NTC | NTC |
B | 6RD4_16S | 6RD4_ITS | 6RD5_16S | 6RD5_ITS | 6RD6_16S | 6RD6_ITS | 6RD7_16S | 6RD7_ITS | 6LVD5_16S | 0.0002 pM Std | 0.0002 pM Std | 0.0002 pM Std |
C | 6RD4_16S | 6RD4_ITS | 6RD5_16S | 6RD5_ITS | 6RD6_16S | 6RD6_ITS | 6RD7_16S | 6RD7_ITS | 6LVD5_16S | 0.002 pM Std | 0.002 pM Std | 0.002 pM Std |
D | 6RD4_16S | 6RD4_ITS | 6RD5_16S | 6RD5_ITS | 6RD6_16S | 6RD6_ITS | 6RD7_16S | 6RD7_ITS | 6LVD5_16S | 0.02 pM Std | 0.02 pM Std | 0.02 pM Std |
E |
0.2 pM Std
0.2 pM Std
F
6RD4_16S | 6RD4_ITS | 6RD5_16S | 6RD5_ITS | 6RD6_16S | 6RD6_ITS | 6RD7_16S | 6RD7_ITS | 6LVD5_ITS | 0.2 pM Std | 0.2 pM Std |
G
0.2 pM Std |
20 pM Std
H
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)
...
110
...
1100
...
2 ul
...
Primer Premix (10X)
...
110
...
220
...
4 ul
...
Ultra Pure Water
...
110
...
440
...
16 ul
...
Total Volume
...
110
...
1760
Add 4 ul of template, pool, or standards to each well:
...
...
1
...
2
...
3
...
4
...
5
...
6
...
7
...
8
...
9
...
10
...
11
...
12
...
A
...
NTC
...
NTC
...
NTC
...
B
...
0.0002 pM Std
...
0.0002 pM Std
...
0.0002 pM Std
...
C
...
0.002 pM Std
...
0.002 pM Std
...
0.002 pM Std
...
D
...
0.02 pM Std
...
0.02 pM Std
...
0.02 pM Std
...
E
...
0.2 pM Std
...
0.2 pM Std
...
0.2 pM Std
...
F
...
2 pM Std
...
2 pM Std
...
2 pM Std
F | 6RD4_16S | 6RD4_ITS | 6RD5_16S | 6RD5_ITS | 6RD6_16S | 6RD6_ITS | 6RD7_16S | 6RD7_ITS | 6LVD5_ITS | 2 pM Std | 2 pM Std | 2 pM Std |
G | 6RD4_16S | 6RD4_ITS | 6RD5_16S | 6RD5_ITS | 6RD6_16S | 6RD6_ITS | 6RD7_16S | 6RD7_ITS | 6LVD5_ITS | 20 pM Std | 20 pM Std | 20 pM Std |
H | 6RD4_16S | 6RD4_ITS | 6RD5_16S | 6RD5_ITS | 6RD6_16S | 6RD6_ITS | 6RD7_16S | 6RD7_ITS | 6LVD5_ITS | NS6_Pool_1:1000 | NS6_Pool_1:1000 | NS6_Pool_1:1000 |
Results:
Plate Name | Average Result (nanomoles) |
|---|---|
6RD4_16S | 70.66 |
6RD4_ITS | 22.2 |
6RD5_16S | 76.68 |
6RD5_ITS | 86.82 |
6RD6_16S | 32.57 |
6RD6_ITS | 45.66 |
6RD7_16S | 45.71 |
6RD7_ITS | 67.98 |
6LVD5_16S | 50.03 |
6LVD5_ITS | 14.98 |
NS6_Pool | 12.17 |
Full qPCR results below:
| View file | ||
|---|---|---|
|