Dandekar & Chandler: Colony PCR for non-E. coli strains: Difference between revisions
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* Scoop up a grain of rice-sized blob of cells (use a stick, blue loops are too big to fit to the bottom of eppi tube) | * Scoop up a grain of rice-sized blob of cells (use a stick, blue loops are too big to fit to the bottom of eppi tube) | ||
* Resuspend in 50 ul sterile dH20 | * Resuspend in 50 ul sterile dH20 | ||
* Boil for 5 min at 100°C - caps may pop open, so use the fasteners or watch for swamping by dirty | * Boil for 5 min at 100°C - caps may pop open, so use the fasteners or watch for swamping by dirty water | ||
* Spin down tubes at 16000g for 5 min | * Spin down tubes at 16000g for 5 min | ||
* Use 5 ul of supernatant in PCR reaction | * Use 5 ul of supernatant in PCR reaction | ||
Revision as of 12:20, 24 November 2014
Back to Protocols
A pseudomonas colony PCR method that seems to work
- Scoop up a grain of rice-sized blob of cells (use a stick, blue loops are too big to fit to the bottom of eppi tube)
- Resuspend in 50 ul sterile dH20
- Boil for 5 min at 100°C - caps may pop open, so use the fasteners or watch for swamping by dirty water
- Spin down tubes at 16000g for 5 min
- Use 5 ul of supernatant in PCR reaction
