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

lacI, placI, pSS3 digestion