IGEM:Peking University/2008/Experiment/PKU 0717 G2 1

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PKU_080717_G2_1


Screen for pGREF505-lexO clonies

Plasmid Miniprep-G4M1278-13~24
Sequencing-G4M1278-14/17/24
Result for last sequencing: G4M1278-1&2 are right point mutations.

Point mutation-T1278G/T1743G
Using right wtGAL4 TG-11
Primer Name:T1743G-F & T1743G-R


PCR system
Volume (μL)
10×buffer iPCR 5
pGREG505/(TEF-lacI-adhlacS-GFP) 5
dNTP 5
KOD Plus 1
Gal4 template 1
T1278G-F-Primer 0.75
T1278G-R-Primer 0.75
ddH2O 36.5


conditions
Temperature(°C) Time Length(0:00:00)
94 02:00
98 00:10 10 cycles
68 00:07 30 cycles
Hold 4°C
98 00:10 25 cycles
68 10:00 25 cycles
Hold 4°C


Result: have about 7000 band, waiting for sequencing check

Digestion-point muation product-overnight/4°C
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