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21/08/13
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Reculturing Bacteria
- The P. putida grew overnight, which proved that we had viable cells.
- The bacteria was then re-streaked to single colonies and left to grow overnight in a 37C incubator.
Isolation of P. putida DNA
- 8 1.5ml samples were isolated from each of the 2 broths that were prepared yesterday.
- The first few steps of CTAB protocol was used to prep cells for DNA isolation;
- The 8 samples were spun down at 10000 rpm for 5 mins.
- The supernatant was discarded.
- cells were then resuspended in 1ml of TE.
- 740ul of this new mixture was transferred to 8 new eppendorf tubes.
- 20ul of lysozyme was added to each of these 8 samples.
- A maxwell robot was then used to complete the DNA extraction process.
Hydrodynamic shearing of Herring Sperm DNA
- A 20G syringe was used ~24 times to squirt the herring sperm DNA in order to make it less viscous.
- 30G syringe was also used
- This method seems to work better than the sonification.
