From 2013.igem.org
{"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"}
"
