Matt:LabNotes/2013-6-26: Difference between revisions
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===Page Size Selection of 0 gap probes=== | ===Page Size Selection of 0 gap probes=== | ||
USER/DpnII digested 0 gap probes: | USER/DpnII digested 0 gap probes: | ||
Noi made 11.36ug/400ul [http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-5-29] | Noi made 11.36ug/~400ul [http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-5-29] | ||
I made 10.58ug/180ul [http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-5-16] | I made 10.58ug/180ul [http://genome-tech.ucsd.edu/LabNotes/index.php/Matt:LabNotes/2013-5-16] | ||
<!--If Combined: 11.36ug + 10.58ug / (410ul + 180ul) = 21.94ug/590ul = 37.19ng/ul (590ul)--> | <!--If Combined: 11.36ug + 10.58ug / (410ul + 180ul) = 21.94ug/590ul = 37.19ng/ul (590ul)--> | ||
To run ~3ug/gel, I use 100ul of probes that Noi made for each of the 4 gels | To run ~3ug/gel, I use ~100ul of probes that Noi made for each of the 4 gels | ||
* Ran TBU gel at 220V for 30 min with samples and ladder prepared below | * Ran TBU gel at 220V for 30 min with samples and ladder prepared below | ||
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| align="center" style="background:#f0f0f0;"|'''4X Volume''' | | align="center" style="background:#f0f0f0;"|'''4X Volume''' | ||
|- | |- | ||
| Enzyme digested probes (~11.36/400ul)|| | | Enzyme digested probes (~11.36/400ul)||106||424 | ||
|- | |- | ||
| TBE-Urea Buffer 2X|| | | TBE-Urea Buffer 2X||106||424 | ||
|- | |- | ||
| H2O||0||0 | | H2O||0||0 | ||
|- | |- | ||
| Total|| | | Total||212||848 | ||
|- | |- | ||
| | | |
Revision as of 18:39, 26 June 2013
Page Size Selection of 0 gap probes
USER/DpnII digested 0 gap probes:
Noi made 11.36ug/~400ul [1] I made 10.58ug/180ul [2]
To run ~3ug/gel, I use ~100ul of probes that Noi made for each of the 4 gels
- Ran TBU gel at 220V for 30 min with samples and ladder prepared below
- Heated TBE buffer in microwave
- Denatured samples/ladder for 10 min at 75C before putting on cold rack for 1 min
- Size selected for brightest band below 200bp
Components | 1X Volume | 4X Volume |
Enzyme digested probes (~11.36/400ul) | 106 | 424 |
TBE-Urea Buffer 2X | 106 | 424 |
H2O | 0 | 0 |
Total | 212 | 848 |
Components | 1X Volume | 4X Volume |
10-bp DNA ladder (1 ug/ul) | 1 | 4 |
TBE-Urea Buffer 2X | 5 | 20 |
H2O | 4 | 16 |
total | 10 | 40 |