Team:UNITN-Trento/Notebook/Labposts/07/19

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{"date":"2013-07-06","author":"viola","title":"<html>cloning of EFE in pSpac and pXyl vectors</html>","content":"<html> Today i start the cloning to insert the ethylene forming enzyme in both the bacillus plasmids. To make this i digested the vectors and also EFE+B0015 in pSB1C3 with EcorI and PstI. {{:Team:UNITN-Trento/Templates/Styles/Spoiler|LIGATION TABLE 1| <html> <center> <table> <tr> <th>LIGATION</th> <th>EFE+B0015 in pSPAC</th> <th>EFE+B0015 in pXYL</th> </tr> <tr> <th>PLASMID CONC.</th> <td>20 µl</td> <td>20 µl</td> </tr> <tr> <th>PLASMID LENGHT</th> <td>7082 BP</td> <td>3594 BP</td> </tr> <tr> <th>INSERT CONCENTRATION</th> <td>20 µl</td> <td>20 µl</td> </tr> <tr> <th>INSERT LENGHT</th> <td>1251 BP</td> <td>1251 BP</td> </tr> <tr> <th>PLASMID USED</th> <td>50 ng/µl</td> <td>50 ng/µl</td> </tr> <tr> <th>REACTION VOLUME</th> <td>20 µl</td> <td>20 µl</td> </tr> <tr> <th>BUFFER</th> <td>10 µl</td> <td>10 µl</td> </tr> </table> </center> </html>}} <html> </html>}} {{:Team:UNITN-Trento/Templates/Styles/Spoiler|LIGATION TABLE PSPAC| <html> <center> <table> <tr> <th><font color=\"red\">EFE IN PSPAC</font></th> <th>CTRL</th> <th>1:1</th> <th>1:2</th> <th>1:4</th> </tr> <tr> <th>INSERT (µl)</th> <td>0</td> <td>0.44</td> <td>0.88</td> <td>1.76</td> </tr> <tr> <th>WATER (µl)</th> <td>14.5 </td> <td>14.06</td> <td>13.62</td> <td>12.74</td> </tr> <tr> <th>LIGASE (µl)</th> <td>1 </td> <td>1 </td> <td>1</td> <td>1</td> </tr> <tr> <th>PLASMID (µl)</th> <td>2.5</td> <td>2.5</td> <td>2.5</td> <td>2.5</td> </tr> <tr> <th>BUFFER (µl)</th> <td>2</td> <td>2</td> <td>2</td> <td>2</td> </tr> </table> </center> </html>}} {{:Team:UNITN-Trento/Templates/Styles/Spoiler|LIGATION TABLE PXYL| <html> <center> <table> <tr> <th><font color=\"red\">EFE IN PXYL</font></th> <th>CTRL</th> <th>1:1</th> <th>1:2</th> <th>1:4</th> </tr> <tr> <th>INSERT (µl)</th> <td>0</td> <td>0.87</td> <td>1.74</td> <td>3.48</td> </tr> <tr> <th>WATER (µl)</th> <td>14.5 </td> <td>13.63</td> <td>12.76</td> <td>11.02</td> </tr> <tr> <th>LIGASE (µl)</th> <td>1 </td> <td>1 </td> <td>1</td> <td>1</td> </tr> <tr> <th>PLASMID (µl)</th> <td>2.5</td> <td>2.5</td> <td>2.5</td> <td>2.5</td> </tr> <tr> <th>BUFFER (µl)</th> <td>2</td> <td>2</td> <td>2</td> <td>2</td> </tr> </table> </center> </html>}} <html> </html>}} <html> </html>}} </html>","tags":"BACILLUS"}
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"date" : "2013-07-10",
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"author" : "fabio",
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"title" : " ripenator v1.0 ",
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"content" : "<html> this morning, Bruno is not available, so I will take care of the ripenator. I have to replace all the cultures with new LB and new bacteria : I’m gonna put 300 ml of LB in each beutas, then connect immediately a beuta with LB and 3ml of not-induced bacteria to the control banana!! After that, I will dissolve 78 ul of MESA in the second beuta and connect to the blocked banana! As far as the ethylene sample is concerned, I have to put 3 ml of bacteria in the last beuta, wait for the OD lecture to reach the value of 0,5 and then induce the culture with Arabinose to produce Ethylene! Finally I can connect it with the mature banana. </html>",
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"tags" : " ripenator-mesa-ethylene "
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}
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Latest revision as of 10:36, 3 October 2013

{"date":"2013-07-06","author":"viola","title":"cloning of EFE in pSpac and pXyl vectors","content":" Today i start the cloning to insert the ethylene forming enzyme in both the bacillus plasmids. To make this i digested the vectors and also EFE+B0015 in pSB1C3 with EcorI and PstI. {{:Team:UNITN-Trento/Templates/Styles/Spoiler|LIGATION TABLE 1|

LIGATION EFE+B0015 in pSPAC EFE+B0015 in pXYL
PLASMID CONC. 20 µl 20 µl
PLASMID LENGHT 7082 BP 3594 BP
INSERT CONCENTRATION 20 µl 20 µl
INSERT LENGHT 1251 BP 1251 BP
PLASMID USED 50 ng/µl 50 ng/µl
REACTION VOLUME 20 µl 20 µl
BUFFER 10 µl 10 µl
}} }}
LIGATION TABLE PSPAC
EFE IN PSPAC CTRL 1:1 1:2 1:4
INSERT (µl) 0 0.44 0.88 1.76
WATER (µl) 14.5 14.06 13.62 12.74
LIGASE (µl) 1 1 1 1
PLASMID (µl) 2.5 2.5 2.5 2.5
BUFFER (µl) 2 2 2 2
LIGATION TABLE PXYL
EFE IN PXYL CTRL 1:1 1:2 1:4
INSERT (µl) 0 0.87 1.74 3.48
WATER (µl) 14.5 13.63 12.76 11.02
LIGASE (µl) 1 1 1 1
PLASMID (µl) 2.5 2.5 2.5 2.5
BUFFER (µl) 2 2 2 2
}} }} </html>","tags":"BACILLUS"}

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