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Team:Lethbridge
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<u>Related Information:</u><br> | <u>Related Information:</u><br> | ||
Endonucleases available | Endonucleases available | ||
| - | <table> | + | <table><table border="3"> |
| - | <tr> | + | <tr><td>Endonuclease</td><td>Optimal Buffer**</td><td>Other Buffers</td></tr> |
| - | <td>Endonuclease</td><td>Optimal Buffer</td><td>Other Buffers</td> | + | <tr><td>EcoRV</td><td>None</td><td>2xT(100%); O,G(50-100%)</td></tr> |
| - | </tr> | + | <tr><td>EcoRI</td><td>Red</td><td>O(100%);R(100%)*;2xT(100%)</td></tr> |
| - | <tr><td> | + | <tr><td>BcuI/SpeI</td><td>Tango</td><td>B(50-100%);G(50-100%)</td></tr> |
| - | </tr> | + | <tr><td>XbaI</td><td>Tango</td><td>B,G,2xT(50-100%)</td></tr> |
| + | <tr><td>PstI</td><td>Orange</td><td>R(100%); B,G,T,2xT(50-100%)</td></tr> | ||
| + | <tr><td>DpnI</td><td>Tango</td><td>B,G(100%): O,R,2xT(50-100%)</td></tr> | ||
| + | </table> | ||
| + | *Star Activity<br> | ||
| + | **Optimal Buffer from Fermentas<br><br> | ||
| + | Use pUC19 plasmid as test, it has cut sites for EcoRI, PstI, XbaI (unsure about BcuI/SpeI, DpnI but will try anyways), and none for EcoRV <br> | ||
| + | <u>Red Buffer:</u> EcoRI, PstI, Control (No Enzyme)<br> | ||
| + | <u>Tango Buffer:</u> BcuI/SpeI, XbaI, DpnI, Control (No Enzyme><br><br> | ||
| + | Set up Master Mixes: | ||
| + | <table><table border="3"> | ||
| + | <tr><td><u>Red MM</u></td><td>per tube (µL)</td><td>Total (µL)</td></tr> | ||
| + | <tr><td>MilliQ H<sub>2</sub>0</td><td>13.75</td><td>55</td></tr> | ||
| + | <tr><td>Red Buffer (10x)</td><td>2</td><td>7</td></tr> | ||
| + | <tr><td>pUC19 (10pg/µL)</td><td>2</td><td>7</td></tr> | ||
| + | <tr><td><b>Total</b></td><td>19.75</td><td>69</td></tr> | ||
| + | </table> | ||
| + | |||
Revision as of 03:31, 14 May 2010
University of Lethbridge IGEM team
- Main
- Team
- Projects
- Notebook
- Meetings
- Parts
- Collaborations
blah balh balh
Our team is based yada yada
- Team Pics
- Tab 3
- Team bios
Team pictures
Content 3
Pics of us
Overview
- Project 1
- Porejct 2
Content 1
Content 2
Objective: Test Restriction Endonucleases for Activity
Related Information:
Endonucleases available
| Endonuclease | Optimal Buffer** | Other Buffers |
| EcoRV | None | 2xT(100%); O,G(50-100%) |
| EcoRI | Red | O(100%);R(100%)*;2xT(100%) |
| BcuI/SpeI | Tango | B(50-100%);G(50-100%) |
| XbaI | Tango | B,G,2xT(50-100%) |
| PstI | Orange | R(100%); B,G,T,2xT(50-100%) |
| DpnI | Tango | B,G(100%): O,R,2xT(50-100%) |
**Optimal Buffer from Fermentas
Use pUC19 plasmid as test, it has cut sites for EcoRI, PstI, XbaI (unsure about BcuI/SpeI, DpnI but will try anyways), and none for EcoRV
Red Buffer: EcoRI, PstI, Control (No Enzyme)
Tango Buffer: BcuI/SpeI, XbaI, DpnI, Control (No Enzyme>
Set up Master Mixes:
| Red MM | per tube (µL) | Total (µL) |
| MilliQ H20 | 13.75 | 55 |
| Red Buffer (10x) | 2 | 7 |
| pUC19 (10pg/µL) | 2 | 7 |
| Total | 19.75 | 69 |
May 13
This is what we did today
This is where our electronic lab book data will go, gel pictures will also go here and here is an example of it:
Content 6
Content 7
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