Difference between revisions of "Team:Westminster/Results"
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<h3> Project Results</h3> | <h3> Project Results</h3> | ||
+ | RESULTS<br> | ||
+ | <b>11/09/15</b><br> | ||
+ | Nanodrop results from plasmid prep<br> | ||
+ | OmcA (1) - 260/280= 1.87 260/230=2.10<br> | ||
+ | TOTAL= 114.5 ng/µl<br> | ||
+ | OmcA (2) - 260/280= 1.88 260/230=2.13<br> | ||
+ | TOTAL= 77.9 ng/µl<br> | ||
+ | CymA (1) - 260/280= 1.86 260/230=2.13<br> | ||
+ | TOTAL= 183.5ng/µl<br> | ||
+ | CymA (2) - 260/280= 1.88 260/230=2.06<br> | ||
+ | TOTAL= 107.1ng/µl<br> | ||
+ | MtrC (1) - 260/280= 1.88 260/230=2.15<br> | ||
+ | TOTAL= 166.7ng/µl<br> | ||
+ | MtrC (2) - 260/280= 1.89 260/230=2.18<br> | ||
+ | TOTAL= 91.1 ng/µl<br> | ||
+ | <b>17/09/15<br> | ||
+ | 2 Gels of PCR products extracted yesterday morning (16/09/15) (tube 1, 2, 3, 4, 5, 6, 7, 8, MtrCAB). Gel was run for 40 mins at 100V. <br> | ||
+ | GEL 1</b><br> | ||
+ | <table border=="1" > | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <td valign="top" ><p><b>Lane 1</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 2</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 3</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 4</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 5</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 6</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 7</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 8</b></p></td> | ||
+ | </tr> | ||
− | <p> | + | <tr> |
+ | <td valign="top" ><p><b>DNA ladder (10</b> <b>µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>MtrCAB (2 µl + 8 µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>Tube 1 (2 µl + 8 µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>Tube 2 (2 µl + 8 µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>Tube 3 (2 µl + 8 µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>Tube 4 (2 µl + 8 µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>DNA ladder (10</b> <b>µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>Empty</b></p></td> | ||
+ | </tr> | ||
− | + | </tbody> | |
− | + | </table> | |
− | + | ||
− | + | ||
− | + | ||
− | </ | + | |
+ | <br><br>Both ladders and MtrCAB were the only lanes with results. <br> | ||
+ | <br><br> <br> <br> <br> <br> <br>image here <br> <br> <br> <br> <br> <br> | ||
− | |||
− | |||
− | < | + | <b>GEL 2 </b><br><table border="1"> |
− | < | + | <tbody> |
− | < | + | <tr> |
− | </ | + | <td valign="top" ><p><b>Lane 1</b></p></td> |
+ | <td valign="top" ><p><b>Lane 2</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 3</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 4</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 5</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 6</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 7</b></p></td> | ||
+ | <td valign="top" ><p><b>Lane 8</b></p></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td valign="top" ><p><b>DNA ladder (10</b> <b>µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>Tube 5 (2 µl + 8 µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>Tube 6 (2 µl + 8 µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>Tube 7 (2 µl + 8 µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>Tube 8 (2 µl + 8 µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>DNA ladder (10</b> <b>µl)</b></p></td> | ||
+ | <td valign="top" ><p><b>Empty</b></p></td> | ||
+ | <td valign="top" ><p><b>Empty</b></p></td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
− | < | + | |
− | <p> | + | <br> |
− | < | + | Lane 3 was the only lane without results. |
− | < | + | <br> <br> <br> <br> <br> <br>image here <br> <br> <br> <br> <br> <br> |
− | < | + | |
− | < | + | |
− | </ | + | <br> |
+ | <b>Plasmid prep</b> <br> | ||
+ | 9 LB bottles were inoculated with amplicillin (100µg/ml) or chlorophenicol (34.5µg/ml) and plasmid pSB1C3 or pSB1A3 from yesterday’s transformation and left to incubate overnight (pSB1C3 =tube 2, 4, 5, 7 and MtrCAB; pSB1A3= tube 1, 3, 6, 8).<br><br> | ||
+ | |||
+ | <table border="1"> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <td valign="top" ><p>Steps</p></td> | ||
+ | <td valign="top" ><p>Tube 1,3,6,8 (Amp)</p></td> | ||
+ | <td valign="top" ><p>Tube 2,5,7 (all Chlr) MtrCAB, </p></td> | ||
+ | <td valign="top" ><p>Tube 4 Chlr</p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p>1. Harvest</p></td> | ||
+ | <td valign="top" ><p>Yes 15mins + extra 5 mins</p></td> | ||
+ | <td valign="top" ><p>Yes 15mins + extra 5 mins</p></td> | ||
+ | <td valign="top" ><p>Yes 15mins + extra 5 mins</p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p>2. Resuspend</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p>3. Lyse</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p>4. Precipitate</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p>5. Bind</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p>6. Wash (optional)</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p>7. Wash and Ethanol Removal</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p>8. Elute</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p>9. Recover</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | <td valign="top" ><p>yes</p></td> | ||
+ | </tr> | ||
+ | |||
+ | </tbody> | ||
+ | </table> | ||
+ | |||
+ | <br> | ||
+ | |||
+ | |||
+ | Tube 3 (Amp) - resuspended with 340 µl rather than 250 µl <br> | ||
+ | Tube 7 (Chlr) - 360 µl resuspension buffer added rather than 250 µl <br> | ||
+ | <b>Nanodrop results of plasmid prep </b><br> | ||
+ | <b>Amp 1</b>- 260/280= 1.98 260/230=2.12 <br> | ||
+ | TOTAL= 83.0ng/µl <br> | ||
+ | <b>Chlr 2</b>- 260/280= 1.89 260/230=2.11 <br> | ||
+ | TOTAL= 151.7ng/µl <br> | ||
+ | <b>Amp 3</b>- 260/280= 1.91 260/230=2.12 <br> | ||
+ | TOTAL= 173.5ng/µl <br> | ||
+ | <b>Chlr 4</b>- 260/280= 1.89 260/230=2.18 <br> | ||
+ | TOTAL= 92.1ng/µl <br> | ||
+ | <b>Chlr 5</b>- 260/280= 1.93 260/230=2.23 <br> | ||
+ | TOTAL= 87.8ng/µl <br> | ||
+ | <b>Amp 6</b>- 260/280= 1.92 260/230=2.15 <br> | ||
+ | TOTAL= 129.5ng/µl <br> | ||
+ | <b>Chlr 7</b>- 260/280= 1.91 260/230=2.21 <br> | ||
+ | TOTAL= 80.4ng/µl <br> | ||
+ | <b>Chlr 8</b>- 260/280= 1.94 260/230=2.11 <br> | ||
+ | TOTAL= 128.4ng/µl <br> | ||
+ | <b>MtrCAB</b>- 260/280= 1.96 260/230=2.28 <br> | ||
+ | TOTAL= 38.3ng/µl <br> <br> | ||
+ | |||
+ | Below is the plate that the plasmid DNA that will be freeze dried <br> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <table border="1"> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <td valign="top" ><p><br /> | ||
+ | </p></td> | ||
+ | <td valign="top" ><p><b>1</b></p></td> | ||
+ | <td valign="top" ><p><b>2</b></p></td> | ||
+ | <td valign="top" ><p><b>3</b></p></td> | ||
+ | <td valign="top" ><p><b>4</b></p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p><b>A</b></p></td> | ||
+ | <td valign="top" ><p>1</p></td> | ||
+ | <td valign="top" ><p>9</p></td> | ||
+ | <td valign="top" ><p>17</p></td> | ||
+ | <td valign="top" ><p>25</p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p><b>B</b></p></td> | ||
+ | <td valign="top" ><p>2</p></td> | ||
+ | <td valign="top" ><p>10</p></td> | ||
+ | <td valign="top" ><p>18</p></td> | ||
+ | <td valign="top" ><p>26</p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p><b>C</b></p></td> | ||
+ | <td valign="top" ><p>3</p></td> | ||
+ | <td valign="top" ><p>11</p></td> | ||
+ | <td valign="top" ><p>19</p></td> | ||
+ | <td valign="top" ><p>27</p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p><b>D</b></p></td> | ||
+ | <td valign="top" ><p>4</p></td> | ||
+ | <td valign="top" ><p>12</p></td> | ||
+ | <td valign="top" ><p>20</p></td> | ||
+ | <td valign="top" ><p><br /> | ||
+ | </p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p><b>E</b></p></td> | ||
+ | <td valign="top" ><p>5</p></td> | ||
+ | <td valign="top" ><p>13</p></td> | ||
+ | <td valign="top" ><p>21</p></td> | ||
+ | <td valign="top" ><p><br /> | ||
+ | </p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p><b>F</b></p></td> | ||
+ | <td valign="top" ><p>6</p></td> | ||
+ | <td valign="top" ><p>14</p></td> | ||
+ | <td valign="top" ><p>22</p></td> | ||
+ | <td valign="top" ><p><br /> | ||
+ | </p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p><b>G</b></p></td> | ||
+ | <td valign="top" ><p>7</p></td> | ||
+ | <td valign="top" ><p>15</p></td> | ||
+ | <td valign="top" ><p>23</p></td> | ||
+ | <td valign="top" ><p><br /> | ||
+ | </p></td> | ||
+ | </tr> | ||
+ | |||
+ | <tr> | ||
+ | <td valign="top" ><p><b>H</b></p></td> | ||
+ | <td valign="top" ><p>8</p></td> | ||
+ | <td valign="top" ><p>16</p></td> | ||
+ | <td valign="top" ><p>24</p></td> | ||
+ | <td valign="top" ><p><br /> | ||
+ | </p></td> | ||
+ | </tr> | ||
+ | |||
+ | </tbody> | ||
+ | </table> | ||
+ | |||
+ | <table><tr><td> | ||
+ | 1. MtrA pSB1C3 <br> | ||
+ | 2. MtrB pSB1C3 <br> | ||
+ | 3. MtrC pSB1C3 <br> | ||
+ | 4. MtrC (1) <br> | ||
+ | 5. MtrC (2) <br> | ||
+ | 6. CymA (1) <br> | ||
+ | 7. CymA (2) </td><td> | ||
+ | 8. OmcA <br> | ||
+ | 9. OmcA (1) <br> | ||
+ | 10. OmcA (2) <br> | ||
+ | 11. MtrA-His <br> | ||
+ | 12. MtrB-His <br> | ||
+ | 13. MtrC-His <br> | ||
+ | 14. CymA-His <br> | ||
+ | 15. OmcA-His | ||
+ | 16- OmcA-CymA(1)</td><td> | ||
+ | 17. OmcA-CymA (2) <br> | ||
+ | 18. MtrCAB <br> | ||
+ | 19. Chlr 2 <br> | ||
+ | 20. Chlr 4 <br> | ||
+ | 21. Chlr 5 <br> | ||
+ | 22. Chlr 7</td></tr></table> | ||
Revision as of 07:12, 18 September 2015
Project Results
RESULTS11/09/15
Nanodrop results from plasmid prep
OmcA (1) - 260/280= 1.87 260/230=2.10
TOTAL= 114.5 ng/µl
OmcA (2) - 260/280= 1.88 260/230=2.13
TOTAL= 77.9 ng/µl
CymA (1) - 260/280= 1.86 260/230=2.13
TOTAL= 183.5ng/µl
CymA (2) - 260/280= 1.88 260/230=2.06
TOTAL= 107.1ng/µl
MtrC (1) - 260/280= 1.88 260/230=2.15
TOTAL= 166.7ng/µl
MtrC (2) - 260/280= 1.89 260/230=2.18
TOTAL= 91.1 ng/µl
17/09/15
2 Gels of PCR products extracted yesterday morning (16/09/15) (tube 1, 2, 3, 4, 5, 6, 7, 8, MtrCAB). Gel was run for 40 mins at 100V.
GEL 1
Lane 1 |
Lane 2 |
Lane 3 |
Lane 4 |
Lane 5 |
Lane 6 |
Lane 7 |
Lane 8 |
DNA ladder (10 µl) |
MtrCAB (2 µl + 8 µl) |
Tube 1 (2 µl + 8 µl) |
Tube 2 (2 µl + 8 µl) |
Tube 3 (2 µl + 8 µl) |
Tube 4 (2 µl + 8 µl) |
DNA ladder (10 µl) |
Empty |
Both ladders and MtrCAB were the only lanes with results.
image here
GEL 2
Lane 1 |
Lane 2 |
Lane 3 |
Lane 4 |
Lane 5 |
Lane 6 |
Lane 7 |
Lane 8 |
DNA ladder (10 µl) |
Tube 5 (2 µl + 8 µl) |
Tube 6 (2 µl + 8 µl) |
Tube 7 (2 µl + 8 µl) |
Tube 8 (2 µl + 8 µl) |
DNA ladder (10 µl) |
Empty |
Empty |
Lane 3 was the only lane without results.
image here
Plasmid prep
9 LB bottles were inoculated with amplicillin (100µg/ml) or chlorophenicol (34.5µg/ml) and plasmid pSB1C3 or pSB1A3 from yesterday’s transformation and left to incubate overnight (pSB1C3 =tube 2, 4, 5, 7 and MtrCAB; pSB1A3= tube 1, 3, 6, 8).
Steps |
Tube 1,3,6,8 (Amp) |
Tube 2,5,7 (all Chlr) MtrCAB, |
Tube 4 Chlr |
1. Harvest |
Yes 15mins + extra 5 mins |
Yes 15mins + extra 5 mins |
Yes 15mins + extra 5 mins |
2. Resuspend |
yes |
yes |
yes |
3. Lyse |
yes |
yes |
yes |
4. Precipitate |
yes |
yes |
yes |
5. Bind |
yes |
yes |
yes |
6. Wash (optional) |
yes |
yes |
yes |
7. Wash and Ethanol Removal |
yes |
yes |
yes |
8. Elute |
yes |
yes |
yes |
9. Recover |
yes |
yes |
yes |
Tube 3 (Amp) - resuspended with 340 µl rather than 250 µl
Tube 7 (Chlr) - 360 µl resuspension buffer added rather than 250 µl
Nanodrop results of plasmid prep
Amp 1- 260/280= 1.98 260/230=2.12
TOTAL= 83.0ng/µl
Chlr 2- 260/280= 1.89 260/230=2.11
TOTAL= 151.7ng/µl
Amp 3- 260/280= 1.91 260/230=2.12
TOTAL= 173.5ng/µl
Chlr 4- 260/280= 1.89 260/230=2.18
TOTAL= 92.1ng/µl
Chlr 5- 260/280= 1.93 260/230=2.23
TOTAL= 87.8ng/µl
Amp 6- 260/280= 1.92 260/230=2.15
TOTAL= 129.5ng/µl
Chlr 7- 260/280= 1.91 260/230=2.21
TOTAL= 80.4ng/µl
Chlr 8- 260/280= 1.94 260/230=2.11
TOTAL= 128.4ng/µl
MtrCAB- 260/280= 1.96 260/230=2.28
TOTAL= 38.3ng/µl
Below is the plate that the plasmid DNA that will be freeze dried
|
1 |
2 |
3 |
4 |
A |
1 |
9 |
17 |
25 |
B |
2 |
10 |
18 |
26 |
C |
3 |
11 |
19 |
27 |
D |
4 |
12 |
20 |
|
E |
5 |
13 |
21 |
|
F |
6 |
14 |
22 |
|
G |
7 |
15 |
23 |
|
H |
8 |
16 |
24 |
|
1. MtrA pSB1C3 2. MtrB pSB1C3 3. MtrC pSB1C3 4. MtrC (1) 5. MtrC (2) 6. CymA (1) 7. CymA (2) |
8. OmcA 9. OmcA (1) 10. OmcA (2) 11. MtrA-His 12. MtrB-His 13. MtrC-His 14. CymA-His 15. OmcA-His 16- OmcA-CymA(1) |
17. OmcA-CymA (2) 18. MtrCAB 19. Chlr 2 20. Chlr 4 21. Chlr 5 22. Chlr 7 |