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

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{"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"}