IGEM:Peking/2007/Switch-Notebook/2007-8-2

T1TE and T1T2
T1TE digestion by ClaI/SalI and T1T2 digestion by NotI/XhoI, then rescued.

LacZa-plx009 miniprep
NdeI and XhoI digestion using H buffer, send to sequencing.

sequencing result
plx008 is right

plx009-5 turned out to be plx003!!!

So actually we have already constructed lacZa-Plx003!

But there is still some problems with enzymatic digestion results.

We will change our construction plan due to this result!

plx006 digestion using XhoI and H buffer
result: plx006 can not be cut by XhoI wihout any known reason.

CD2, RD3, RS3, CD3 PCR and rescue
using relevant primers to PCR and then rescue by gel.

==plx007/8 transform JM110/DH5a

T1TE and T1T2 ligate with pcc010
4 degrees in fridge overnight with a concentration ratio of 4:1 between fragment and vector