"
21/08/13
From 2013.igem.org
(Difference between revisions)
AliceHaworth (Talk | contribs) |
AliceHaworth (Talk | contribs) (→hydrodynamic shearing of Herring Sperm DNA) |
||
| (2 intermediate revisions not shown) | |||
| Line 15: | Line 15: | ||
*The ''P. putida'' grew overnight, which proved that we had viable cells. | *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. | *The bacteria was then re-streaked to single colonies and left to grow overnight in a 37C incubator. | ||
| - | ==Isolation of ''P. putida''DNA == | + | ==Isolation of ''P. putida'' DNA == |
*8 1.5ml samples were isolated from each of the 2 broths that were prepared yesterday. | *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 first few steps of CTAB protocol was used to prep cells for DNA isolation; | ||
| Line 24: | Line 24: | ||
*20ul of lysozyme was added to each of these 8 samples. | *20ul of lysozyme was added to each of these 8 samples. | ||
* A maxwell robot was then used to complete the DNA extraction process. | * 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 | + | *30G syringe was also used |
| - | *This method | + | *This method seems to work better than the sonification. |
Latest revision as of 13:55, 22 August 2013
| Home | Team | Official Team Profile | Project | Parts Submitted to the Registry | Modeling | Notebook | Safety | Attributions |
|---|
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.
