IGEM:Peking/2007/Switch-Notebook/2007-8-29
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Preparation of liquid medium LB and LB Amp
miniprep and digestion test
sulA, RS2, RD3-plx012 plasmid is isolated.
Double digestion system:
H buffer 2uL
plasmid 7uL
XhoI/SalI 0.25uL each
ddH2O 10.5uL
colony PCR of LacIq-T
many colonies are right!
ligation of CD2 with plx012
re-digestion of CD1-plx012
plasmid 7uL
buffer H 2uL
XhoI/NdeI 0.5uL each
H2O 10uL to a total 20uL system
result: right!
sequencing of LacIq-pcc009
No signal. There must be a problem with either our plasmid or the primer!