Alice:LabNotes/2007-12-7
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Experiment 1: PCR on newly ordered PGP1Ver3 primer
The new primer PGP1Ver 3 came in with 24 different forward and reverse primers each (200uM). Then the primers are diluted to 10uM by mixing with 10uL Forward + 10uL Reverse + 180 uL H2O for experimental purpose and to preserve the original samples.
The primers are then ready for PCR. The reagents used are listed below:
' | one reaction | total |
Template | 1 | 72 |
2X Master Mix | 25 | 1800 |
10uM forward primer + reverse primer | 1 | 72 |
H2O | 23 | 1656 |
95C 3min -> 35 cycles of (95C 15S -> 58C 30S -> 72C 1min) -> 72C 3min
Notes: The templates used are: GM20431 (row A-B), GC1F (row C-D), GC1EP (row E-F)
Also, since the cap of the PCR machine wasn't properly secured, so few wells have evaporated. Most of them are fine to use, but well F9 is almost empty, so experiments on these wells will be repeated again later.