Team:Tokyo Metropolitan/Notebook/A30
From 2011.igem.org
(Difference between revisions)
(→-VisualizatioLink titlen-) |
(→-Visualization-) |
||
(3 intermediate revisions not shown) | |||
Line 1: | Line 1: | ||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
{{:Team:Tokyo_Metropolitan/Header}} | {{:Team:Tokyo_Metropolitan/Header}} | ||
<html> | <html> | ||
<div style="width: 900px; margin-left: 20px; padding-top: 25px; padding-left: 20px;"> | <div style="width: 900px; margin-left: 20px; padding-top: 25px; padding-left: 20px;"> | ||
</html> | </html> | ||
+ | = -Speeding Up- = | ||
+ | Amplified HNS(wt),I20260 with PCR and purified these PCR products. | ||
+ | |||
= -Visualization- = | = -Visualization- = | ||
Plasmid extraction and purification to get J23100・I714051,but there was enough plasmid. | 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) |
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)