-Speeding Up-

Amplified HNS(wt),I20260 with PCR and purified these PCR products.

-Visualization-

Plasmid extraction and purification to get J23100・I714051,but there was enough plasmid.

@plasmid extraction OriTr

@DNA refine OriTr

@DNA densitometry

@restriction emzyme(EcoRⅠ and SpeⅠ, ScoRⅠ and XbaⅠ) OriTr，OriTr+GFP

-Targeting-

-DNA refining

P0440

no.1 + no.3 143

no.4 + no.5 130

J23100

no.1 + no.3 170

no.4 + no.5 208

K317037

no.3 + no.5 210

-Digestion

P0440xba+pst

J23100Spe+pst

K317037xba+pst

after digested, incubated for 5 hours at 37 deg C.

-Electrophoresis

Results

Only had cut K317037

-Gel extraction

Gel extracted for K317037, then DNA refined

-Mesurement of concentration

J23100 -> 90

K317037 no.2 102, no.4 75

(ng/ul)