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Revision as of 16:17, 12 September 2010 by
Imb10
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PCR Result
9/6 ~ 9/9 PCR Result
PCR composition
Materials
Amount
Template
100pmole
Primer A
50pmole
Primer B
50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)
2ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)
0.25ul
10x buffer (10x cloned Pfu Reaction buffer)
5ul
DW (3rd sterile water)
rest
Total
50ul
Exp 1
Segment
Annealing Temperature
Result
a
STAT1
53*C
-
-
b
V_STAT1
53*C
-
-
c
Fusion Antibody receptor
53*C
-
-
d
Signal peptide
56*C
-
-
e
Ig-like
56*C
-
-
f
Signal Peptide + Ig-like
56*C
-
-
g
Antibody
56*C
-
-
h
V_Fusion Antibody Receptor
53*C
-
-
i
B_FGFR1
56*C
-
-
j
VB_FGFR1
56*C
-
-
Exp 2
Segment
Annealing Temperature
Result
a
STAT1
48*C
60%
2139bp
b
V_STAT1
48*C
-
-
Exp 3
Segment
Annealing Temperature
Result
a
STAT1
50*C
100%
2139bp
b
V_STAT1
50*C
-
-
Exp 4
Segment
Annealing Temperature
Result
a
Fusion Antibody receptor
50*C
-
-
b
Signal peptide
50*C
-
-
c
Ig-like
50*C
60%
339bp
d
Signal Peptide + Ig-like
50*C
-
-
e
Antibody
50*C
-
-
f
V_Fusion Antibody Receptor
50*C
-
-
9/10 ~ 9/12 PCR Result
PCR composition
Materials
Amount
Template
150pmole
Primer A
50pmole
Primer B
50pmole
dNTP (dNTP mixture 2.5mM TaKaRa)
5ul
Taq (Pfu Turbo® DNA Polymerase 2.5U/ μl STRATAGENE)
0.25ul
10x buffer (10x cloned Pfu Reaction buffer)
5ul
DW (3rd sterile water)
rest
Total
50ul
Exp 1
Segment
Annealing Temperature
Result
a
STAT1
60*C
-
-
b
V_STAT1
60*C
-
-
c
Fusion Antibody receptor
60*C
70%
1095bp
d
Signal peptide
60*C
-
-
e
Ig-like
60*C
60%
339bp
f
Signal Peptide + Ig-like
60*C
-
-
g
Antibody
60*C
100%
669bp
h
V_Fusion Antibody Receptor
60*C
-
-
i
B_FGFR1
60*C
-
-
j
VB_FGFR1
60*C
-
-
x1
negative 1
60*C
-
-
x2
negative 2
60*C
-
-
Exp 2
Segment
Annealing Temperature
Result
a
STAT1
58*C
60%
2139bp
b
V_STAT1
58*C
-
-
c
Fusion Antibody receptor
58*C
80%
1095bp
d
Signal peptide
58*C
80%
87bp
e
Ig-like
58*C
100%
339bp
f
Signal Peptide + Ig-like
58*C
80%
426bp
g
Antibody
58*C
100%
669bp
h
V_Fusion Antibody Receptor
58*C
-
-
i
B_FGFR1
58*C
-
-
j
VB_FGFR1
58*C
-
-
x
negative
58*C
-
-
Exp 3
Segment
Annealing Temperature
Result
a
STAT1
62*C
-
-
b
V_STAT1
62*C
-
-
c
Fusion Antibody receptor
62*C
10%
1095bp
d
Signal peptide
62*C
-
-
e
Ig-like
62*C
-
-
f
Signal Peptide + Ig-like
62*C
-
-
g
Antibody</font><td width = "200px">62*C
<td width = "223px">-</td><td width = "70px">-</td>
<tr align=center> <td width = "40px">h</td><td width = "255px">V_Fusion Antibody Receptor</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td> </tr> <tr align=center> <td width = "40px">i</td><td width = "255px">B_FGFR1</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td> </tr> <tr align=center> <td width = "40px">j</td><td width = "255px">VB_FGFR1</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td> </tr> <tr align=center> <td width = "40px">x</td><td width = "255px">negative</td><td width = "200px">62*C</td><td width = "223px">-</td><td width = "70px">-</td> </tr> </table>
Exp 4
Segment
Annealing Temperature
Result
a
STAT1
56*C
-
-
b
V_STAT1
56*C
-
-
c
Fusion Antibody receptor
56*C
5%
1095bp
d
Signal peptide
56*C
-
-
e
Ig-like
56*C
80
339bp
f
Signal Peptide + Ig-like
56*C
-
-
g
Antibody
56*C
-
-
h
V_Fusion Antibody Receptor
56*C
-
-
i
B_FGFR1
56*C
-
-
j
VB_FGFR1
56*C
-
-
x
negative
56*C
-
-
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