"
Exeter/9 August 2013
From 2013.igem.org
Digestions of CMY
| Culture | NanoDrop (ng/ul) | ul to get 250ng |
|---|---|---|
| Magenta K592012 | 106.9 | 2.34 |
| Yellow K592010 | 57.1 | 4.38 |
| Yellow S05058 | 67.1 | 3.73 |
| RFP control | 42.9 | 5.83 |
| AMP plasmid | 82.5 | 3.03 |
| B0015 | 52.1 | 4.80 |
Xul DNA
2.5ul NEB2
0.5ul BSA (make sure diluted)
0.5ul of each required enzyme
(make up to 16ul with no-nuclease water)
Working out water needed:
Magenta = 2.34 + x = 16 x = 13.66ul
Yellow K592010 = 4.38 + x = 16 x = 11.62ul
Yellow S05058 = 3.73 + x = 16 x = 12.27ul
RFP = 5.83 + x = 16 x = 10.17ul
AMP plasmid = 3.03 + x = 16 x = 12.97ul
B0015 = 4.80 + x = 16 x = 11.20ul
Gel
| Lane number | Content |
|---|---|
| 1 | Hyperladder 1kb |
| 2 | Yellow (S05058) |
| 3 | Magenta |
| 4 | Yellow (K592010) |
| 5 | RFP |
| 6 | AMP plasmid |
| 7 | B0015 |
| 8 | Hyperladder 1kb |
Gel has had limited success. Obviously, parts like [http://parts.igem.org/Part:BBa_B0015 B0015] are too small to see on this gel, as they run much further than the ladder can indicate. Yellow and RFP are the only clear bands on the gel.
Ligations
We have replicates of each, so there is:
Yellow ([http://parts.igem.org/Part:BBa_S05058 S05058]) + Terminator #1
Yellow ([http://parts.igem.org/Part:BBa_S05058 S05058]) + Terminator #2
Magenta + Terminator #1
Magenta + Terminator #2
Yellow ([http://parts.igem.org/Part:BBa_K592010 K592010]) + Terminator #1
Yellow ([http://parts.igem.org/Part:BBa_K592010 K592010]) + Terminator #2
RFP + Terminator #1
RFP + Terminator #2
Take me back to the notebook.
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