Team:KAIST-Korea/Project/Methods/PCR Result

From 2010.igem.org

 



PCR Experiment

9/6 ~ 9/9 PCR Result

PCR composition

MaterialsAmount
Template100pmole
Primer A50pmole
Primer B50pmole
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
Total50ul



Exp 1

SegmentAnnealing TemperatureResult
aSTAT153*C--
bV_STAT153*C--
cFusion Antibody receptor53*C--
dSignal peptide56*C--
eIg-like56*C--
fSignal Peptide + Ig-like56*C--
gAntibody56*C--
hV_Fusion Antibody Receptor53*C--
iB_FGFR156*C--
jVB_FGFR156*C--


KAIST-exp1.png



Exp 2

SegmentAnnealing TemperatureResult
aSTAT148*C60%2139bp
bV_STAT148*C--


KAIST-exp2.png



Exp 3

SegmentAnnealing TemperatureResult
aSTAT150*C100%2139bp
bV_STAT150*C--


KAIST-exp3.png



Exp 4

SegmentAnnealing TemperatureResult
aFusion Antibody receptor50*C--
bSignal peptide50*C--
cIg-like50*C60%339bp
dSignal Peptide + Ig-like50*C--
eAntibody50*C--
fV_Fusion Antibody Receptor50*C--


KAIST-exp4.png



9/10 ~ 9/12 PCR Result

PCR composition

MaterialsAmount
Template150pmole
Primer A50pmole
Primer B50pmole
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
Total50ul



Exp 1

SegmentAnnealing TemperatureResult
aSTAT160*C--
bV_STAT160*C--
cFusion Antibody receptor60*C70%1095bp
dSignal peptide60*C--
eIg-like60*C60%339bp
fSignal Peptide + Ig-like60*C--
gAntibody60*C100%669bp
hV_Fusion Antibody Receptor60*C--
iB_FGFR160*C--
jVB_FGFR160*C--
x1negative 160*C--
x2negative 260*C--


KAIST-exp5.png



Exp 2

SegmentAnnealing TemperatureResult
aSTAT158*C60%2139bp
bV_STAT158*C--
cFusion Antibody receptor58*C80%1095bp
dSignal peptide58*C80%87bp
eIg-like58*C100%339bp
fSignal Peptide + Ig-like58*C80%426bp
gAntibody58*C100%669bp
hV_Fusion Antibody Receptor58*C--
iB_FGFR158*C--
jVB_FGFR158*C--
xnegative58*C--


KAIST-exp6.png



Exp 3

SegmentAnnealing TemperatureResult
aSTAT162*C--
bV_STAT162*C--
cFusion Antibody receptor62*C10%1095bp
dSignal peptide62*C--
eIg-like62*C--
fSignal Peptide + Ig-like62*C--
gAntibody62*C--
hV_Fusion Antibody Receptor62*C--
iB_FGFR162*C--
jVB_FGFR162*C--
xnegative62*C--


KAIST-exp7.png



Exp 4

SegmentAnnealing TemperatureResult
aSTAT156*C--
bV_STAT156*C--
cFusion Antibody receptor56*C5%1095bp
dSignal peptide56*C--
eIg-like56*C80%339bp
fSignal Peptide + Ig-like56*C--
gAntibody56*C80%669bp
hV_Fusion Antibody Receptor56*C--
iB_FGFR156*C--
jVB_FGFR156*C--
xnegative56*C--


KAIST-exp8.png



Exp 5

SegmentAnnealing TemperatureResult
aV_STAT164*C--
bB_FGFR164*C5%1398bp
cVB_FGFR164*C--


KAIST-exp9.png



Exp 6

SegmentAnnealing TemperatureResult
aV_STAT164*C--
bB_FGFR164*C3%1398bp
cVB_FGFR164*C--


KAIST-exp10.png



Exp 7

SegmentAnnealing TemperatureResult
aV_STAT168*C--
bB_FGFR168*C10%1398bp
cVB_FGFR168*C--


KAIST-exp12.png



Exp 8

SegmentAnnealing TemperatureResult
aV_STAT170*C--
bB_FGFR170*C3%1398bp
cVB_FGFR170*C--
c5'UTR70*C--
c3'UTR70*C--


KAIST-exp13.png



Exp 9

SegmentAnnealing TemperatureResult
aV_STAT167*C--
bB_FGFR167*C6%1398bp
cVB_FGFR167*C--


KAIST-exp14.png



9/13 ~ 9/16 PCR Result

PCR composition

MaterialsAmount
Template100pmole
Primer A50pmole
Primer B50pmole
Bioneer Accupower HF PCR premix < 1ul
DW (3rd sterile water)rest
Total50ul



Exp 1

SegmentAnnealing TemperatureResult
aSTAT150*C100%2139bp
bSTAT152*C100%2139bp
cSTAT154*C100%2139bp
dSTAT156*C100%2139bp
eSTAT158*C100%2139bp
fSTAT160*C100%2139bp
gV_STAT150*C100%2151bp
hV_STAT152*C100%2151bp
iV_STAT154*C100%2151bp
jV_STAT156*C100%2151bp
kV_STAT158*C100%2151bp
lV_STAT160*C100%2151bp


KAIST-exp21.png



Exp 2

SegmentAnnealing TemperatureResult
aB_FGFR50*C100%1398bp
bB_FGFR52*C100%1398bp
cB_FGFR54*C100%1398bp
dB_FGFR56*C100%1398bp
eB_FGFR58*C100%1398bp
fB_FGFR60*C100%1398bp
gVB_FGFR50*C100%1420bp
hVB_FGFR52*C100%1420bp
iVB_FGFR54*C100%1420bp
jVB_FGFR56*C100%1420bp
kVB_FGFR58*C100%1420bp
lVB_FGFR60*C100%1420bp


KAIST-exp22.png



Exp 3

SegmentAnnealing TemperatureResult
aSignal peptide61*C30%87bp
bIg-like61*C100%339bp
cSignal peptide + Ig-like61*C100%426bp
dSignal peptide59*C30%87bp
eIg-like59*C100%339bp
fSignal peptide + Ig-like59*C100%426bp
g3'UTR52*C50%444bp
h3'UTR54*C50%444bp
i3'UTR56*C100%444bp
j3'UTR58*C100%444bp
k3'UTR60*C100%444bp
l5'UTR52*C100%500bp
m5'UTR54*C100%500bp
n5'UTR56*C100%500bp
o5'UTR58*C100%500bp
p5'UTR69*C70%500bp


KAIST-exp23.png



Exp 4

SegmentAnnealing TemperatureResult
aF_Fusion antibody receptor60*C100%1095bp
bF_Fusion antibody receptor58*C100%1095bp
cAntibody60*C100%669bp
dAntibody58*C100%669bp
eVF_Fusion antibody receptor60*C100%1107bp
fVF_Fusion antibody receptor58*C100%1107bp


KAIST-exp24.png



9/17 PCR result

PCR composition

MaterialsAmount
Template100pmole
Primer A50pmole
Primer B50pmole
Bioneer Accupower HF PCR premix < 1ul
DW (3rd sterile water)rest
Total50ul



Exp 1

SegmentAnnealing TemperatureResult
aSelection marker(41)50*C0%--
bSelection marker(41)52*C0%--
cSelection marker(41)54*C0%--
dSelection marker(41)56*C0%--
eSelection marker(42)50*C0%--
fSelection marker(42)52*C0%--
gSelection marker(42)54*C0%--
hSelection marker(42)56*C0%--
iF_Fusion antibody receptor50*C50%1107bp
jF_Fusion antibody receptor52*C80%1107bp
kB_Fusion antibody receptor54*C100%1398bp
lB_Fusion antibody receptor56*C100%1398bp


KAIST-exp25.png



Exp 2

SegmentAnnealing TemperatureResult
aHA150*C100%87bp
bHA152*C100%87bp
cHA154*C100%87bp
dHA156*C100%87bp
eHA250*C100%87bp
fHA252*C100%87bp
gHA254*C100%87bp
hHA256*C100%87bp
iProximal promoter50*C100%120bp
jProximal promoter52*C100%120bp
kProximal promoter54*C100%120bp
lProximal promoter56*C100%120bp
m5' UTR54*C100%500bp
n5' UTR54*C100%500bp
o3' UTR54*C100%444bp
p3' UTR54*C100%444bp


KAIST-exp26.png



9/18 PCR result

PCR composition

MaterialsAmount
Template100pmole
Primer A50pmole
Primer B50pmole
Bioneer Accupower HF PCR premix < 1ul
DW (3rd sterile water)rest
Total50ul



Exp 1

SegmentAnnealing TemperatureResult
aHA2+3'UTR58*C100%531bp
bF_Fusion antibody receptor58*C100%1107bp
cF_Fusion antibody receptor58*C100%1107bp
dB_Fusion antibody receptor58*C100%1398bp
eB_Fusion antibody receptor58*C100%1398bp


360px



9/19 PCR result

PCR composition

MaterialsAmount
Template100pmole
Primer A50pmole
Primer B50pmole
Bioneer Accupower HF PCR premix < 1ul
DW (3rd sterile water)rest
Total50ul



Exp 1

SegmentAnnealing TemperatureResult
aFusion antibody receptor58*C50%2493bp


360px



9/21 PCR result

PCR composition

MaterialsAmount
Template100pmole
Primer A50pmole
Primer B50pmole
Bioneer Accupower HF PCR premix < 1ul
DW (3rd sterile water)rest
Total50ul



Exp 1

SegmentAnnealing TemperatureResult
aF_Fusion antibody receptor58*C100%1107bp
bB_Fusion antibody receptor58*C100%1398bp
cCore promoter58*C0%--
dSelection marker(42)58*C0%--


360px



Exp 2

SegmentAnnealing TemperatureResult
aSelection marker(41)58*C0%--
bSelection marker(42)58*C0%--


360px



10/5 PCR result

PCR composition

MaterialsAmount
Template100pmole
Primer A50pmole
Primer B50pmole
Bioneer Accupower HF PCR premix < 1ul
DW (3rd sterile water)rest
Total50ul



Exp 1

SegmentAnnealing TemperatureResult
aIg-like58*C100%339bp
bSig + Ig-like58*C100%426bp
cAntibody58*C90%669bp
dF_FGFR58*C50%1095bp
eB_FGFR58*C100%1398bp
fSTAT158*C100%2139bp

KAIST-exp31.png