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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!