Test samples were previously extracted.
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ul/rxn | Reagent | # of rxns | ul needed |
|---|---|---|---|
3 | 5X Kapa HiFi Buffer | 11080 | 330240 |
0.45 | 10M dNTPs | 11080 | 49.536 |
0.3 | Kapa HiFi HotStart DNA Pol | 11080 | 3324 |
7.25 | HPLC H2O | 11080 | 797.5580 |
11 | Total Volume | 11080 | 1210880 |
Add 11 ul to each well of a hard shell, full skirt plate. Add 2 uL of primers and 2uL of template to each well.
Primers: _______________ (fill in when primers arrive).
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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 | P1_A | P2_A | P3_A | P4_A | S1_A | S2_A | S3_A | S4_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 |
|
|
|
|
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