06/17/15
Minpreps & Digests
- Minipreps
- Sigma GenElute kit, elute w/ 50 μL sterilized water.
- Digests
- Check with E/S digests; cannot use PstI because PLrigid & PLrigid4 contain PstI sites
Reagent
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Volume
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Expected: 1. DT009 = 2668, 252 Empty PLflex = 2668, 95 2. DT010 = 2656, 432 Empty PLflex4 = 2656, 274 3. DT011 = 2668, 252 Empty PLrigid = 2668, 95 4. DT012 = 2656, 432 Empty PLrigid4 = 2656, 275
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15 μL/lane; 1% agarose; Ladder
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DNA(plasmid) |
2.0 μL
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10X buffer |
1.5
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EcoRI |
1.0
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SpeI |
1.0
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dH2O |
9.5
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15 μL --> 37°C/ ~15 min.
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CONCLUSIONS:
- DT009 - success.
- DT010 - success.
- DT011 - success.
- DT012 - success.
- Next step:
- Sequence verify all clones (extra precaution; linker part is small)
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