Haynes Lab:Notebook/CRISPR Editing/2015/07/28: Difference between revisions
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| Reagent || Vol. μL || (x12) | | Reagent || Vol. μL || (x12) | ||
|- | |- | ||
| DNA (20 ng) || | | DNA (20 ng) || 0.5 || 6.0 | ||
|- | |- | ||
| PCR H<sub>2</sub>O || 4. | | PCR H<sub>2</sub>O || 4.0 || 48.0 | ||
|- | |- | ||
| || 4.5 || 54.0 | | || 4.5 || 54.0 | ||
|} | |} | ||
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* Template mix, 13.5 into each rep 1 well (A1, A4, A7...) | |||
* Primer mix, 31.5 into each rep 1 well (A1, B1, C1...H1...) | |||
PCR - Bio-Rad CFX96 | PCR - Bio-Rad CFX96 | ||
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* 40x [95°C, 10 sec / 57°C, 10 sec / 72°C, 10 sec (measure)] | * 40x [95°C, 10 sec / 57°C, 10 sec / 72°C, 10 sec (measure)] | ||
* 72°C, 30 sec | * 72°C, 30 sec | ||
* Melt: 70°C/ 5 sec -- +0.1°C (measure) --> 95°C | * Melt: 70°C/ 5 sec -- +0.1°C (measure) --> 95°C/ 5 sec | ||
Revision as of 06:10, 29 July 2015
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07/28/2015
mCherry qPCR/melt pre-course test (at CSH)
Note: reference primers (GAPDH B2) are not here yet
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PCR - Bio-Rad CFX96
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