IGEM:Peking University/2008/Experiment/PKU 0621 G2 1

TITLE: PCR-pPT1-Gal4-Phusion 50ul*4

	Volume (μL)
Template	0.5
dNTP	2
Enzyme	0.5
Buffer	10
F-Primer	0.2
R-Primer	0.2
ddH2O	36.6

conditions

Temperature	Time Length(0:00:00)
98°C	01:30
98°C	00:10	30 cycles
60.6,63.1,64.5,68.2°C	00:30	30 cycles
72°C	01:30	30 cycles
72°C	07:00
4°C	Hold

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TITLE: PCR-Budding Yeast Genome-Gal4-Phusion 50ul*4

	Volume (μL)
Template	0.5
dNTP	2
Enzyme	0.5
Buffer	10
F-Primer	0.2
R-Primer	0.2
ddH2O	36.6

conditions

Temperature	Time Length(0:00:00)
98°C	01:30
98°C	00:10	30 cycles
60.6,63.1,64.5,68.2°C	00:30	30 cycles
72°C	01:30	30 cycles
72°C	07:00
4°C	Hold

---

TITLE: PCR-pPT-Gal4-Transtart 20ul*6

	Volume (μL)
Template	0.5
dNTP	2
Enzyme	0.5
Buffer	2
F-Primer	0.2
R-Primer	0.2
ddH2O	14

conditions

Temperature	Time Length(0:00:00)
94°C	02:00
94°C	01:00	4 cycles
50°C	02:00	4 cycles
72°C	05:00	4 cycles
94°C	01:00	30 cycles
55.2,55.6,57.6,59.5,61.3,63.9°C	01:00	30 cycles
72°C	03:00	30 cycles
72°C	10:00
4°C	Hold

---

Ligation-pEasy-T1/Gal4-E.Coli-Amp⁺,K⁺(BlueWhiteSelection)

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