IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-10-23: Difference between revisions
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Very carcinogenic :D 40uL EtBr, 20min | Very carcinogenic :D 40uL EtBr, 20min | ||
{| | |||
[[image: 102306_zsprecut.jpg|thumb| | |- | ||
| [[image: 102306_zsprecut.jpg|left|thumb|350px|see above for legend]] | |||
[[image: 102306_zspostcut.jpg|thumb| | | [[image: 102306_zspostcut.jpg|left|thumb|350px|verification correct band cutout]] | ||
|} | |||
===gel purification and pcr purification=== | ===gel purification and pcr purification=== | ||
Line 37: | Line 37: | ||
==sequencing== | ==sequencing== | ||
Sent 4 orders: 11 Vf2&vr and 22 vf2&vr, rush order. | 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. |
Latest revision as of 22:04, 28 October 2006
Redo of yesterday's procedure
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.