IGEM:Peking/2007/Count-Conjugation-Notebook/2007-8-21

Tandem OriT by Qu Mingzhi

Ligation: PlacI-I741051 <- E0240 & R0010 <- E0240 (III)

• Ligate the E0240 fragment and PlacI-I741051 vector .
• Ligate the E0240 fragment and R0010 vector .
• use super fast T4 DNA ligase
• super fast T4 DNA ligase Ligation system contains:

2   µl     vector
3   µl     E0240 fragment 
0.5 µl     ''super fast T4 DNA ligase''
1   µl     10X T4 ligase buffer
3.5 µl     dH20
--------------------------
10 µl     Total

transformation PlacI-I741051 <- E0240 & R0010 <- E0240(III)

NEXT DAY: Received clones.