IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-10-23

CIP of digested 011/sp
1h@37C

gel of post-CIP'd
1h15@130V. 1.2%


 * 1: 1kb+
 * 2: undigested 11
 * 3: empty
 * 4: 11/sp marker
 * 5: 11/sp (not shown)
 * 6: 11/sp (not shown)
 * 7: 11/sp (not shown)
 * 8: 11/sp (not shown)
 * 9: 11/sp marker
 * 10: 1kb+

post-stain
Very carcinogenic :D 40uL EtBr, 20min

gel purification and pcr purification
2000mg gel; purified into 6 tubes. sent all 6 tubes into 1 pcr purification. eluted in 30 eb.

ligation

 * J36011 /SP + J36010 /XP (latest), respectively.
 * BB vector: 70ng @ (70 ng/uL) = 1 uL
 * BB insert: 3*(~1800/3700bp)*70ng=102.16 ng @ (20.2 ng/uL) = 5.06 uL
 * 5x dilution: 2uL
 * h20: 1.94uL

sequencing
Sent 4 orders: 11 Vf2&vr and 22 vf2&vr, rush order.

Transformation
Transformants plated at 8:00 PM, placed in incubator at 8:25 PM.