IGEM:PKU Beijing/2009/Notebook/AND Gate 1/Input/2009/07/12/Shuke Wu

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Every plate is very well: more than 100 clones
PCR
14 tubes: L1×3+L2×3+t2×3+t3×3 and 2 negative controls
Master mix 5ul each, primer (standard primer) 0.5uL each, template;

Gel electrophoresis
Products of PCR
Marker: 100bp 250bp 500bp 750bp 1kb 2kb 3kb 5kb
loading buffer and DNA dye: 6×
Voltage and time: 60V 5min; 120V 15min

Lane 1: t2-1;
Lane 2~4: t3-3~1;
Lane 5: t2-3;
Lane 7: t2-1+2;
Lane 8: negative control1;
Lane 9: marker;
Lane 10: negative control2;
Lane 11~13: L2-3~1;
Lane 16~18: L1-3~1;

Result
There is a polluted line at about 1kb place, but it did not confuse us. The right place of L1 & L2 is about 1.3kb and of t2 & t3 is about 800bp.
L1-1~3, L2-1~3, t2-1&3 and t3-1~2 should be the positive clones.
4 clones were successfully constructed:
L1 & L2: 2×B0034-C0012-B0015
T2 & t3: 2×B0034-C0040-B0015

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