Test Plates
BS_PLT1 _Norm (4 samples)
BS_PLT2_Norm (4 samples)
Well C1 Normalized to 10ng/ul
PCR MasterMix
ul/rxn | Reagent | # of rxns | ul needed |
|---|---|---|---|
3 | 5X Kapa HiFi Buffer | 80 | 240 |
0.45 | 10M dNTPs | 80 | 36 |
0.3 | Kapa HiFi HotStart DNA Pol | 80 | 24 |
7.25 | HPLC H2O | 80 | 580 |
11 | Total Volume | 80 | 880 |
...
Make 1:1000 dilutions of column 1,2,3 and 6,7,8 from the PCR plates by adding 1 ul to 999 ul TE in a deep well plate: (Will fill in with proper sample names.)
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|
A | S1_A | S2_A | S3_A | S4_A | S5_A | S6_A | S7_A | S8_A |
| NTC | NTC | NTC |
B |
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| 0.0002 pM Std | 0.0002 pM Std | 0.0002 pM Std |
C |
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| 0.002 pM Std | 0.002 pM Std | 0.002 pM Std |
D |
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| 0.02 pM Std | 0.02 pM Std | 0.02 pM Std |
E |
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| 0.2 pM Std | 0.2 pM Std | 0.2 pM Std |
F |
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| 2 pM Std | 2 pM Std | 2 pM Std |
G |
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| 20 pM Std | 20 pM Std | 20 pM Std |
H |
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...
Average results:
Temp | Avg uM |
|---|---|
63 | 6.95 |
61 | 9.64 |
59 | 10.36 |
57 | 10.9 |
55 | 10.29 |
53 | 10.46 |
50 | 10.22 |
48 | 8.99 |