IGEM:Peking/2007/Switch-Notebook/2007-8-13
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inactivate T1TE-pcc010
using phenol/choloroform to inactivate NotI/XhoI enzyme
rescue by precipitation
T1T2-pcc010 digestion test
using SacII and T buffer with BSA in it
re-PCR lac and SS3
lac-T
primer: lacO3-XhoI-XbaI-F and LacO1-SalI-SpeI-R
SS3-T
primer: SS3-XhoI-XbaI-F and SS3-SalI-SpeI-R
electrophoresis result to show that we have got both fragment finally!
gel rescue!
re-colony PCR of T1TE-plx010
still not right!
digestion of T1TE-T1T2-pcc010
using SacII
PCR CD1, CD2, RS1-3 and RD1-3
using different templates and primers for different fragments
result: slight bands for all the fragments, we will see the final result after rescue
==single digestion test for pcc010 using ClaI or SalI(both in H buffer) to test its enzymatic sites
PCR lacIq using KOD
buffer 5uL
dNTP 5uL
Mg 2uL
p-f/p-r 1uL each
KOD 1uL
ddH2O 35uL
template toothpick with some colony
94 degrees for 15s, 68 degrees for 30s and 72 degrees for 20s, 30 cycles