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From 2010.igem.org

Lab work

Restriction digestion of plasmid backbone pSB1T3 (~1,0 μg) with restriction enzymes EcoRI and PstI:

pSB1T3	2,5 μL
Buffer H (Roche)	2,0 μL
EcoRI (1 U/μg)	1,0 μL
PstI (1 U/μg)	1,0 μL
H2O	13,5 μL

The digested pSB1T3 was cut from gel and isolate the band. However, the gel extraction kit didn’t work as well as we had hoped, it turned out to be the DNA disappearing kit. Tomorrow a new day.