Team:Tokyo Metropolitan/Notebook/A30

From 2011.igem.org

(Difference between revisions)
(→Speeding Up)
(→-Visualization-)
 
(5 intermediate revisions not shown)
Line 1: Line 1:
 +
{{:Team:Tokyo_Metropolitan/Header}}
 +
<html>
 +
<div style="width: 900px; margin-left: 20px; padding-top: 25px; padding-left: 20px;">
 +
</html>
= -Speeding Up- =
= -Speeding Up- =
Amplified HNS(wt),I20260 with PCR and purified these PCR products.
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)

Retrieved from "http://2011.igem.org/Team:Tokyo_Metropolitan/Notebook/A30"