Team:Tokyo Metropolitan/Notebook/A30
From 2011.igem.org
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@restriction emzyme(EcoRⅠ and SpeⅠ, ScoRⅠ and XbaⅠ) OriTr,OriTr+GFP | @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) |
Latest revision as of 15:39, 5 October 2011
-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)